Hybrid tomato plant named HMX8148

ABSTRACT

A hybrid tomato plant, designated HMX8148 is disclosed. The invention relates to the seeds of hybrid tomato designated HMX8148, to the plants and plant parts of hybrid tomato designated HMX8148, and to methods for producing a tomato plant by crossing the hybrid tomato HMX8148 with itself or another tomato plant.

FIELD OF THE INVENTION

The present invention relates to the field of agriculture, to new anddistinctive hybrid tomato plants, such as a hybrid plant designatedHMX8148 and to methods of making and using such hybrids.

BACKGROUND OF THE INVENTION

The following description includes information that may be useful inunderstanding the present invention. It is not an admission that any ofthe information provided herein is prior art or relevant to thepresently claimed inventions, or that any publication specifically orimplicitly referenced is prior art.

Tomato is an important and valuable vegetable crop. Thus, a continuinggoal of plant breeders is to develop stable, high yielding tomatohybrids that are agronomically sound or unique. The reasons for thisgoal are to maximize the amount of fruit produced on the land used(yield) as well as to improve the fruit appearance, the fruit shape andsize, eating and processing qualities and/or the plant agronomic andhorticultural qualities. To accomplish this goal, the tomato breedermust select and develop tomato plants that have the traits that resultin superior parental lines that combine to produce superior hybrids.

SUMMARY OF THE INVENTION

The following embodiments and aspects thereof are described inconjunction with systems, tools and methods which are meant to beexemplary, not limiting in scope.

In various embodiments, one or more of the above-described problems havebeen reduced or eliminated, while other embodiments are directed toother improvements.

According to the invention, in some embodiments there is provided anovel hybrid tomato designated HMX8148, also interchangeably referred toas ‘hybrid tomato HMX8148’, ‘tomato hybrid HMX8148’ or ‘HMX8148’.

This invention thus relates to the seeds of hybrid tomato designatedHMX8148, to the plants or parts of hybrid tomato designated HMX8148, toplants or parts thereof comprising all the physiological andmorphological characteristics of hybrid tomato designated HMX8148 orparts thereof, and/or having all the physiological and morphologicalcharacteristics of hybrid tomato designated HMX8148, and/or having oneor more of or all the characteristics of hybrid tomato designatedHMX8148 listed in Table 1 including but not limited to as determined atthe 5% significance level when grown in the same environmentalconditions, and/or having one or more of the physiological andmorphological characteristics of hybrid tomato designated HMX8148 listedin Table 1 including but not limited to as determined at the 5%significance level when grown in the same environmental conditionsand/or having all the physiological and morphological characteristics ofhybrid tomato designated HMX8148 listed in Table 1 including but notlimited to as determined at the 5% significance level when grown in thesame environmental conditions and/or having one or more of thephysiological and morphological characteristics of hybrid tomatodesignated HMX8148 listed in Table 1 when grown in the sameenvironmental conditions and/or having all the physiological andmorphological characteristics of hybrid tomato designated HMX8148 listedin Table 1 when grown in the same environmental conditions. Theinvention also relates to variants, mutants and trivial modifications ofthe seed or plant of hybrid tomato designated HMX8148.

Plant parts of the hybrid tomato plant designated HMX8148 of the presentinvention are also provided, such as, but not limited to, a scion, arootstock, a fruit, a leaf, a flower, a peduncle, a stalk, a root, astamen, an anther, a pistil, a pollen or an ovule obtained from thehybrid plant. The present invention provides fruit of the hybrid tomatoplant designated HMX8148 of the present invention. Such fruit and partsthereof could be used as fresh products for consumption or in processesresulting in processed products such as food products comprising one ormore harvested parts of the hybrid tomato designated HMX8148, such asprepared fruit or parts thereof, canned fruit or parts thereof,freeze-dried or frozen fruits or parts thereof, diced fruits, juices,prepared fruit cuts, canned tomatoes, pastes, sauces, purees, catsupsand the like. All such products are part of the present invention andthe like. The harvested parts or food products can be or can comprisehybrid tomato fruit from hybrid tomato designated HMX8148. The foodproducts might have undergone one or more processing steps such as, butnot limited to cutting, washing, mixing, frizzing, canning, etc. Allsuch products are part of the present invention. The present inventionalso provides plant parts or cells of the hybrid tomato plant designatedHMX8148, wherein a plant regenerated from said plants parts or cells hasone or more of, or all the phenotypic and morphological characteristicsof hybrid tomato designated HMX8148, such as one or more of or all thecharacteristics of hybrid tomato plant designated HMX8148, listed inTable 1 including but not limited to as determined at the 5%significance level when grown in the same environmental conditions. Allsuch parts and cells are part of the present invention.

The plants and seeds of the present invention include those that may beof an essentially derived variety as defined in section 41(3) of thePlant Variety Protection Act of The United States of America, e.g., avariety that is predominantly derived from hybrid tomato designatedHMX8148 or from a variety that i) is predominantly derived from hybridtomato designated HMX8148, while retaining the expression of theessential characteristics that result from the genotype or combinationof genotypes of hybrid tomato designated HMX8148; ii) is clearlydistinguishable from hybrid tomato designated HMX8148; and iii) exceptfor differences that result from the act of derivation, conforms to theinitial variety in the expression of the essential characteristics thatresult from the genotype or combination of genotypes of the hybridtomato plant designated HMX8148.

In another aspect, the present invention provides regenerable cells. Insome embodiments, the regenerable cells are for use in tissue culture ofhybrid tomato designated HMX8148. In some embodiments, the tissueculture is capable of regenerating plants comprising all thephysiological and morphological characteristics of hybrid tomatodesignated HMX8148, and/or having all the physiological andmorphological characteristics of hybrid tomato designated HMX8148,and/or having one or more of the physiological and morphologicalcharacteristics of hybrid tomato designated HMX8148, and/or having thecharacteristics of hybrid tomato designated HMX8148. In someembodiments, the regenerated plants have the characteristics of hybridtomato designated HMX8148 listed in Table 1 including but not limited toas determined at the 5% significance level when grown in the sameenvironmental conditions and/or have all the physiological andmorphological characteristics of hybrid tomato designated HMX8148 listedin Table 1 including but not limited to as determined at the 5%significance level when grown in the same environmental conditionsand/or have one or more of the physiological and morphologicalcharacteristics hybrid tomato designated HMX8148 listed in Table 1including but not limited to as determined at the 5% significance levelwhen grown in the same environmental conditions and/or have all thephysiological and morphological characteristics of hybrid tomatodesignated HMX8148 listed in Table 1 when grown in the sameenvironmental conditions.

In some embodiments, the plant parts and cells used to produce suchtissue cultures will be embryos, meristematic cells, seeds, callus,pollens, leaves, anthers, pistils, stamens, roots, root tips, stems,petioles, fruits, cotyledons, hypocotyls, ovaries, seed coats, fruits,stalks, endosperms, flowers, axillary buds or the like. Protoplastsproduced from such tissue culture are also included in the presentinvention. The tomato leaves, shoots, roots and whole plants regeneratedfrom the tissue culture, as well as the fruits produced by saidregenerated plants are also part of the invention. In some embodiments,the whole plants regenerated from the tissue culture have one, more thanone, or all the physiological and morphological characteristics oftomato hybrid designated HMX8148 listed in Table 1, including but notlimited to as determined at the 5% significance level when grown in thesame environmental conditions.

The invention also discloses methods for vegetatively propagating aplant of the present invention. In the present application, vegetativelypropagating can be interchangeably used with vegetative reproduction. Insome embodiments, the methods comprise collecting parts of a hybridtomato designated HMX8148 and regenerating a plant from said parts. Insome embodiments, one of the parts can be for example a stem. In someembodiments, the methods can be for example a stem cutting that isrooted into an appropriate medium according to techniques known by theone skilled in the art. Plants and parts thereof, including but notlimited to fruits thereof, produced by such methods are also included inthe present invention. In another aspect, the plants and parts thereofsuch as stems and fruits thereof produced by such methods comprise allthe physiological and morphological characteristics of hybrid tomatodesignated HMX8148, and/or have all the physiological and morphologicalcharacteristics of hybrid tomato designated HMX8148 and/or have thephysiological and morphological characteristics of hybrid tomatodesignated HMX8148 and/or have one or more of the characteristics ofhybrid tomato designated HMX8148. In some embodiments, plants, parts orfruits thereof produced by such methods consist of one, more than one,or all the physiological and morphological characteristics of tomatohybrid designated HMX8148 listed in Table 1, including but not limitedto as determined at the 5% significance level when grown in the sameenvironmental conditions.

Further included in the invention are methods for producing fruitsand/or seeds from the hybrid tomato designated HMX8148. In someembodiments, the methods comprise growing a hybrid tomato designatedHMX8148 to produce tomato fruits and/or seeds. In some embodiments, themethods further comprise harvesting the hybrid tomato fruits and/orseeds. Such fruits and/or seeds are parts of the present invention. Insome embodiments, such fruits and/or seeds have all the physiologicaland morphological characteristics of the fruits and/or seeds of hybridtomato designated HMX8148 (e.g. those listed in Table 1) when grown inthe same environmental conditions and/or have one or more of thephysiological and morphological characteristics of the fruits and/orseeds of the hybrid tomato designated HMX8148 (e.g. those listed inTable 1) when grown in the same environmental conditions and/or have thecharacteristics of the fruits and/or seeds of the hybrid tomatodesignated HMX8148 (e.g. those listed in Table 1) when grown in the sameenvironmental conditions.

Also included in this invention are methods for producing a tomatoplant. In some embodiments, the tomato plant is produced by crossing thehybrid tomato designated HMX8148 with itself or other tomato plant. Insome embodiments, the other plant can be a hybrid tomato other than thehybrid tomato designated HMX8148. In other embodiments, the other plantcan be a tomato inbred line. When crossed with an inbred line, in someembodiments, a “three-way cross” is produced. When crossed with itself(i.e. when a tomato HMX8148 is crossed with another hybrid tomatoHMX8148 plant or when self-pollinated), or with another, differenthybrid tomato, in some embodiments, a “four-way” cross is produced. Suchthree and four-way hybrid seeds and plants produced by growing saidthree and four-way hybrid seeds are included in the present invention.Methods for producing a three and four-way hybrid tomato seedscomprising (a) crossing hybrid tomato designated HMX8148 tomato plantwith a different tomato inbred line or hybrid and (b) harvesting theresultant hybrid tomato seed are also part of the invention. The hybridtomato seeds produced by the method comprising crossing hybrid tomatodesignated HMX8148 tomato plant with a different tomato plant such as atomato inbred line or hybrid, and harvesting the resultant hybrid tomatoseed are included in the invention, as are included the hybrid tomatoplant or parts thereof and seeds produced by said grown hybrid tomatoplants.

Further included in the invention are methods for producing tomato seedsand plants made thereof. In some embodiments, the methods compriseself-pollinating the hybrid tomato designated HMX8148 and harvesting theresultant hybrid seeds. Tomato seeds produced by such method are alsopart of the invention.

In another embodiment, this invention relates to methods for producing ahybrid tomato designated HMX8148 from a collection of seeds.

In some embodiments, the collection contains both seeds of inbred parentline(s) of hybrid tomato designated HMX8148 seeds and hybrid seeds ofHMX8148. Such a collection of seeds might be a commercial bag of seeds.In some embodiments, said methods comprise planting the collection ofseeds. When planted, the collection of seeds will produce inbred parentlines of hybrid tomato HMX8148 and hybrid plants from the hybrid seedsof HMX8148. In some embodiments, said inbred parent lines of hybridtomato designated HMX8148 plants are identified as having a decreasedvigor compared to the other plants (i.e. hybrid plants) grown from thecollection of seeds. In some embodiments, said decreased vigor is due tothe inbreeding depression effect and can be identified for example by aless vigorous appearance for vegetative and/or reproductivecharacteristics including a shorter plant height, small fruit size,fruit shape, fruit color or other characteristics. In some embodiments,seeds of the inbred parent lines of the hybrid tomato HMX8148 arecollected and, if new inbred parent plants thereof are grown and crossedin a controlled manner with each other, the hybrid tomato HMX8148 willbe recreated.

This invention also relates to methods for producing other tomato plantsderived from hybrid tomato HMX8148 and to the tomato plants derived bythe use of methods described herein.

In some embodiments, such methods for producing a tomato plant derivedfrom hybrid tomato HMX8148 comprise (a) self-pollinating the hybridtomato HMX8148 plant at least once to produce a progeny plant derivedfrom the hybrid tomato HMX8148. In some embodiments, the methods furthercomprise (b) crossing the progeny plant derived from the hybrid tomatoHMX8148 with itself or a second tomato plant to produce a seed of aprogeny plant of a subsequent generation. In some embodiments, themethods further comprise (c) growing the progeny plant of the subsequentgeneration. In some embodiments, the methods further comprise (d)crossing the progeny plant of the subsequent generation with itself or asecond tomato plant to produce a tomato plant further derived from thehybrid tomato HMX8148. In further embodiments, steps (b), step (c)and/or step (d) are repeated for at least 1, 2, 3, 4, 5, 6, 7, 8, ormore generations to produce a tomato plant derived from the hybridtomato HMX8148. In some embodiments, within each crossing cycle, thesecond plant is the same plant as the second plant in the last crossingcycle. In some embodiments, within each crossing cycle, the second plantis different from the second plant in the last crossing cycle.

Another method for producing a tomato plant derived from hybrid tomatoHMX8148, comprises (a) crossing the hybrid tomato HMX8148 plant with asecond tomato plant to produce a progeny plant derived from the hybridtomato HMX8148. In some embodiments, the method further comprises (b)crossing the progeny plant derived from the hybrid tomato HMX8148 withitself or a second tomato plant to produce a seed of a progeny plant ofa subsequent generation. In some embodiments, the method furthercomprises (c) growing the progeny plant of the subsequent generation. Insome embodiments, the method further comprises (d) crossing the progenyplant of the subsequent generation with itself or a second tomato plantto produce a tomato plant derived from the hybrid tomato HMX8148. In afurther embodiment, steps (b), (c) and/or (d) are repeated for at least1, 2, 3, 4, 5, 6, 7, 8, or more generations to produce a tomato plantderived from the hybrid tomato HMX8148. In some embodiments, within eachcrossing cycle, the second plant is the same plant as the second plantin the last crossing cycle. In some embodiments, within each crossingcycle, the second plant is different from the second plant in the lastcrossing cycle.

In one aspect, the present invention provides methods of introducing asingle locus conversion conferring one or more desired trait(s) into thehybrid tomato HMX8148, and plants, fruits and/or seeds obtained fromsuch methods. In another aspect, the present invention provides methodsof modifying a single locus conversion conferring one or more desiredtrait(s) into the hybrid tomato HMX8148, and plants, fruits and/or seedsobtained from such methods. The desired trait(s) may be, but notexclusively, a single gene. In some embodiments, the gene is a dominantallele. In some embodiments, the gene is a partially dominant allele. Insome embodiments, the gene is a recessive allele. In some embodiments,the gene or genes will confer such traits, including but not limited tomale sterility, herbicide resistance, insect resistance, resistance forbacterial, fungal, mycoplasma or viral disease, enhanced plant qualitysuch as improved drought or salt tolerance, water-stress tolerance,improved standability, enhanced plant vigor, improved shelf life,delayed senescence or controlled ripening, enhanced nutritional qualitysuch as increased sugar content or increased sweetness, increasedtexture, flavor and aroma, improved fruit length and/or size, protectionfor color, fruit shape, uniformity, length or diameter, refinement ordepth, lodging resistance, yield and recovery, improve fresh cutapplication, specific aromatic compounds, specific volatiles, fleshtexture and specific nutritional components. For the present inventionand the skilled artisan, disease is understood to include, but notlimited to fungal diseases, viral diseases, bacterial diseases,mycoplasma diseases, or other plant pathogenic diseases and a diseaseresistant plant will encompass a plant resistant to fungal, viral,bacterial, mycoplasma, and other plant pathogens. In one aspect, thegene or genes may be naturally occurring tomato gene(s) and/orspontaneous or induced mutations(s). In another aspect, genes aremutated, modified, genetically engineered through the use of NewBreeding Techniques described herein. In some embodiments, the methodfor introducing the desired trait(s) is a backcrossing process by makinguse of a series of backcrosses to at least one of the parent lines ofhybrid tomato designated HMX8148 (a.k.a. hybrid tomato HMX8148 or tomatohybrid HMX8148) during which the desired trait(s) is maintained byselection. At least one of the parent lines of hybrid tomato designatedHMX8148 possesses the desired trait(s) by the backcrossing process, andthe desired trait(s) is inherited by the hybrid tomato progeny plants byconventional breeding techniques known to breeders of ordinary skill inthe art. The single gene converted plants or single locus convertedplants that can be obtained by the methods are included in the presentinvention.

When dealing with a gene that has been modified, for example through NewBreeding Techniques, the trait (genetic modification) could be directlymodified into the newly developed hybrid tomato plant and/or at leastone of the parent lines of hybrid tomato HMX8148. Alternatively, if thetrait is not modified into each newly developed hybrid tomato plantand/or at least one of the parent lines of hybrid tomato HMX8148,another typical method used by breeders of ordinary skill in the art toincorporate the modified gene is to take a line already carrying themodified gene and to use such line as a donor line to transfer themodified gene into the newly developed hybrid tomato plant and/or atleast one of the parent lines of the newly developed hybrid. The samewould apply for a naturally occurring trait or one arising fromspontaneous or induced mutations.

In some embodiments, the backcross breeding process of hybrid tomatoHMX8148 comprises (a) crossing one of the parental inbred line plants ofhybrid tomato HMX8148 with plants of another line that comprise thedesired trait(s) to produce F1 progeny plants. In some embodiments, theprocess further comprises (b) selecting the F1 progeny plants that havethe desired trait(s). In some embodiments, the process further comprises(c) crossing the selected F1 progeny plants with the parental inbredtomato lines of hybrid tomato HMX8148 plants to produce backcrossprogeny plants. In some embodiments, the process further comprises (d)selecting for backcross progeny plants that have the desired trait(s)and essentially all the physiological and morphological characteristicsof the tomato parental inbred line of hybrid tomato HMX8148 to produceselected backcross progeny plants. In some embodiments, the processfurther comprises (e) repeating steps (c) and (d) one, two, three, four,five six, seven, eight, nine or more times in succession to produceselected, second, third, fourth, fifth, sixth, seventh, eighth, ninth orhigher backcross progeny plants that have the desired trait(s) andessentially all the characteristics of the parental inbred tomato lineof hybrid tomato HMX8148, and/or have the desired trait(s) andessentially all the physiological and morphological characteristics ofthe parental tomato inbred line of hybrid tomato HMX8148, and/or havethe desired trait(s) and otherwise essentially all the physiological andmorphological characteristics of the parental inbred tomato line oftomato hybrid HMX8148, including but not limited to when grown in thesame environmental conditions or including but not limited to at a 5%significance level when grown in the same environmental conditions. Thetomato plants or seed produced by the methods are also part of theinvention, as are the hybrid tomato HMX8148 plants that comprised thedesired trait. Backcrossing breeding methods, well known to one skilledin the art of plant breeding will be further developed in subsequentparts of the specification.

An embodiment of this invention is a method of making a backcrossconversion of hybrid tomato HMX8148. In some embodiments, the methodcomprises crossing one of the parental tomato inbred line plants ofhybrid tomato HMX8148 with a donor plant comprising a mutant gene(s), anaturally occurring gene(s) or a gene(s) and/or sequence(s) modifiedthrough New Breeding Techniques conferring one or more desired traits toproduce F1 progeny plants. In some embodiments, the method furthercomprises selecting an F1 progeny plant comprising the naturallyoccurring gene(s), mutant gene(s) or gene(s) and/or sequences(s)modified through New Breeding Techniques conferring the one or moredesired traits. In some embodiments, the method further comprisesbackcrossing the selected progeny plant to the parental tomato inbredline plants of hybrid tomato HMX8148. This method may further comprisethe step of obtaining a molecular marker profile of the parental tomatoinbred line plants of hybrid tomato HMX8148 and using the molecularmarker profile to select for the progeny plant with the desired traitand the molecular marker profile of the parental tomato inbred lineplants of hybrid tomato HMX8148. In some embodiments, this methodfurther comprises crossing the backcross progeny plant of the parentaltomato inbred line plant of hybrid tomato HMX8148 containing thenaturally occurring gene(s), the mutant gene(s) or the modified gene(s)and or sequences modified through New Breeding Techniques conferring theone or more desired trait with the second parental inbred tomato lineplants of hybrid tomato HMX8148 in order to produce the hybrid tomatoHMX8148 comprising the naturally occurring gene(s), the mutant gene(s)or modified gene(s) and/or sequences modified through New BreedingTechniques conferring the one or more desired traits. The plants orparts thereof produced by such methods are also part of the presentinvention.

In some embodiments of the invention, the number of loci that may betransferred and/or backcrossed into the parental tomato inbred line ofhybrid tomato HMX8148 is at least 1, 2, 3, 4, 5, or more.

A single locus may contain several genes. A single locus conversion alsoallows for making one or more site specific changes to the plant genome,such as, without limitation, one or more nucleotide changes, deletions,insertions, substitutions, etc. In some embodiments, the single locusconversion is performed by genome editing, a.k.a. genome editing withengineered nucleases (GEEN). In some embodiments, the genome editingcomprises using one or more engineered nucleases. In some embodiments,the engineered nucleases include, but are not limited to Zinc fingernucleases (ZFNs), Transcription Activator-Like Effector Nucleases(TALENs), the CRISPR/Cas system, engineered meganuclease, re-engineeredhoming endonucleases and endonucleases for DNA guided genome editing(Gao et al., Nature Biotechnology (2016), doi: 10.1038/nbt.3547). Insome embodiments, the single locus conversion changes one or severalnucleotides of the plant genome. Such genome editing techniques are someof the techniques now known by the person skilled in the art and hereinare collectively referred to as “New Breeding Techniques”. In someembodiments, one or more above-mentioned genome editing method isdirectly applied on a plant of the present invention, rather than on theparental tomato inbred lines of hybrid tomato HMX8148. Accordingly, acell containing edited genome, or a plant part containing such cell canbe isolated and used to regenerate a novel plant which has a new traitconferred by said genome editing, and essentially all the physiologicaland morphological characteristics of hybrid tomato plant HMX8148.

The invention further provides methods for developing tomato plants in atomato plant breeding program using plant breeding techniques includingbut not limited to, recurrent selection, backcrossing, pedigreebreeding, genomic selection, molecular marker (Isozyme Electrophoresis,Restriction Fragment Length Polymorphisms (RFLPs), Randomly AmplifiedPolymorphic DNAs (RAPDs), Arbitrarily Primed Polymerase Chain Reactions(AP-PCRs), DNA Amplification Fingerprintings (DAFs), SequenceCharacterized Amplified Regions (SCARs), Amplified Fragment LengthPolymorphisms (AFLPs), and Simple Sequence Repeats (SSRs) which are alsoreferred to as Microsatellites, Single Nucleotide Polymorphisms (SNPs),enhanced selection, genetic markers, enhanced selection andtransformation. Seeds, tomato plants, and parts thereof produced by suchbreeding methods are also part of the invention.

The invention also relates to variants, mutants and trivialmodifications of the seed or plant of the hybrid tomato HMX8148 orinbred parental lines thereof. Variants, mutants and trivialmodifications of the seed or plant of hybrid tomato HMX8148 or inbredparental lines thereof can be generated by methods available to oneskilled in the art, including but not limited to, mutagenesis (e.g.,chemical mutagenesis, radiation mutagenesis, transposon mutagenesis,insertional mutagenesis, signature tagged mutagenesis, site-directedmutagenesis, and natural mutagenesis), knock-outs/knock-ins, antisenseoligonucleotides, RNA interference and other techniques such as the NewBreeding Techniques. For more information of mutagenesis in plants, suchas agents or protocols, see Acquaah et al. (Principles of plant geneticsand breeding, Wiley-Blackwell, 2007, ISBN 1405136464, 9781405136464,which is herein incorporated by reference in its entity).

The invention also relates to a mutagenized population of the hybridtomato HMX8148 and methods of using such populations. In someembodiments, the mutagenized population can be used in screening for newtomato plants which comprise essentially one or more of or all themorphological and physiological characteristics of hybrid tomatoHMX8148. In some embodiments, the new tomato plants obtained from thescreening process comprise essentially all the morphological andphysiological characteristics of the hybrid tomato HMX8148, and one ormore additional or different morphological and physiologicalcharacteristics that the hybrid tomato HMX8148 does not have.

This invention is also directed to methods for producing a tomato plantby crossing a first parent tomato plant with a second parent tomatoplant, wherein either the first or second parent tomato plant is ahybrid tomato plant of HMX8148. Further, both first and second parenttomato plants can come from the hybrid tomato plant HMX8148. Further,the hybrid tomato plant HMX8148 can be self-pollinated i.e. the pollenof a hybrid tomato plant HMX8148 can pollinate the ovule of the samehybrid tomato plant HMX8148. When crossed with another tomato plant, ahybrid seed is produced. Such methods of hybridization andself-pollination are well known to those skilled in the art of breeding.

An inbred tomato line such as one of the parental lines of hybrid tomatoHMX8148 has been produced through several cycles of self-pollination andis therefore to be considered as a homozygous line. An inbred line canalso be produced though the dihaploid system which involves doubling thechromosomes from a haploid plant or embryo thus resulting in an inbredline that is genetically stable (homozygous) and can be reproducedwithout altering the inbred line. Haploid plants could be obtained fromhaploid embryos that might be produced from microspores, pollen, anthercultures or ovary cultures or spontaneous haploidy. The haploid embryosmay then be doubled by chemical treatments such as by colchicine or bedoubled autonomously. The haploid embryos may also be grown into haploidplants and treated to induce the chromosome doubling. In either case,fertile homozygous plants are obtained. A hybrid variety is classicallycreated through the fertilization of an ovule from an inbred parentalline by the pollen of another, different inbred parental line. Due tothe homozygous state of the inbred line, the produced gametes carry acopy of each parental chromosome. As both the ovule and the pollen bringa copy of the arrangement and organization of the genes present in theparental lines, the genome of each parental line is present in theresulting F1 hybrid, theoretically in the arrangement and organizationcreated by the plant breeder in the original parental line.

As long as the homozygosity of the parental lines is maintained, theresulting hybrid cross shall be stable. The F1 hybrid is then acombination of phenotypic characteristics issued from two arrangementand organization of genes, both created by a person skilled in the artthrough the breeding process.

Still further, this invention is also directed to methods for producinga tomato plant derived from hybrid tomato HMX8148 by crossing hybridtomato plant HMX8148 with a second tomato plant. In some embodiments,the methods further comprise obtaining a progeny seed from the cross. Insome embodiments, the methods further comprise growing the progeny seed,and possibly repeating the crossing and growing steps with the hybridtomato plant HMX8148 derived plant from 0 to 7 or more times. Thus, anysuch methods using the hybrid tomato plant HMX8148 are part of thisinvention: selfing, backcrosses, hybrid production, crosses topopulations, and the like. All plants produced using hybrid tomato plantHMX8148 as a parent are within the scope of this invention, includingplants derived from hybrid tomato plant HMX8148. In some embodiments,such plants have one, more than one or all the physiological andmorphological characteristics of the hybrid tomato plant HMX8148 listedin Table 1 including but not limited to as determined at the 5%significance level when grown in the same environmental conditions. Insome embodiments, such plants might exhibit additional and desiredcharacteristics or traits such as high seed yield, high seedgermination, seedling vigor, early maturity, high fruit yield, ease offruit setting, disease tolerance or resistance, lodging resistance, andadaptability for soil and climate conditions. Consumer-driven traits,such as a preference for a given fruit size, fruit shape, fruit color,fruit texture, fruit taste, fruit firmness, fruit sugar content areother traits that may be incorporated into new tomato plants developedby this invention.

A tomato plant can also be propagated vegetatively. A part of the plant,for example a shoot tissue, is collected, and a new plant is obtainedfrom the part. Such part typically comprises an apical meristem of theplant. The collected part is transferred to a medium allowingdevelopment of a plantlet, including for example rooting or developmentof shoots, or is grafted onto a tomato plant or a rootstock prepared tosupport growth of shoot tissue. This is achieved using methods wellknown in the art. Accordingly, in one embodiment, a method ofvegetatively propagating a plant of the present invention comprisescollecting a part of a plant according to the present invention, e.g. ashoot tissue, and obtaining a plantlet from said part. In oneembodiment, a method of vegetatively propagating a plant of the presentinvention comprises: (a) collecting tissue of a plant of the presentinvention; (b) rooting said proliferated shoots to obtain rootedplantlets. In one embodiment, a method of vegetatively propagating aplant of the present invention comprises: (a) collecting tissue of aplant of the present invention; (b) cultivating said tissue to obtainproliferated shoots; (c) rooting said proliferated shoots to obtainrooted plantlets. In one embodiment, such method further comprisesgrowing a plant from said plantlets. In one embodiment, a fruit isharvested from said plant. In one embodiments, such fruits and plantshave all the physiological and morphological characteristics of fruitsand plants of hybrid tomato designated HMX8148 when grown in the sameenvironmental conditions. In one embodiment, the fruit is processed intoproducts such as canned tomato fruits and/or parts thereof, juices,freeze dried or frozen fruit and/or parts thereof, fresh or preparedfruit and/or parts thereof or pastes, sauces, purees, catsups and thelike.

The invention is also directed to the use of the hybrid tomato plantHMX8148 in a grafting process. In one embodiment, the hybrid tomatoplant HMX8148 is used as the scion while in another embodiment, thehybrid tomato plant HMX8148 is used as a rootstock.

In some embodiments, the present invention teaches a seed of hybridtomato designated HMX8148, wherein a representative sample of seed ofsaid hybrid is deposited under NCIMB No.43896.

In some embodiments, the present invention teaches a tomato plant, or apart thereof, produced by growing the deposited HMX8148 seed.

In some embodiments, the present invention teaches a tomato plant part,wherein the tomato part is selected from the group consisting of: aleaf, a flower, a fruit, a stalk, a root, a rootstock, a seed, anembryo, a peduncle, a stamen, an anther, a pistil, an ovule, a pollen, acell, a rootstock, and a scion.

In some embodiments, the present invention teaches a tomato plant, or apart thereof, having all the characteristics of hybrid tomato HMX8148 aslisted in Table 1 of this invention including but not limited to asdetermined at the 5% significance level when grown in the sameenvironmental conditions.

In some embodiments, the present invention teaches a tomato plant, or apart thereof, having all the physiological and morphologicalcharacteristics of hybrid tomato HMX8148, wherein a representativesample of seed of said hybrid was deposited under NCIMB No. 43896.

In some embodiments, the present invention teaches a tissue culture ofregenerable cells produced from the plant or part grown from thedeposited HMX8148 seed, wherein cells of the tissue culture are producedfrom a plant part selected from the group consisting of protoplasts,embryos, meristematic cells, callus, pollens, ovules, flowers, seeds,leaves, roots, root tips, anthers, stems, petioles, fruits, axillarybuds, cotyledons and hypocotyls. In some embodiments, the plant partincludes protoplasts produced from a plant grown from the depositedHMX8148 seed.

In some embodiments, the present invention teaches a compositioncomprising regenerable cells produced from the plant or part thereofgrown from the deposited hybrid HMX8148 seed, or other part or cellthereof. In some embodiments, the composition comprises a growth media.In some embodiments, the growth media is solid or a syntheticcultivation medium. In some embodiments, the composition is a tomatoplant regenerated from the tissue culture from a plant grown from thedeposited HMX8148 seed, said plant having the characteristics of hybridtomato HMX8148, wherein a representative sample of seed of said hybridis deposited under NCIMB No. 43896.

In some embodiments, the present invention teaches a tomato fruitproduced from the plant grown from the deposited HMX8148 seed.

In some embodiments, such fruits have all the physiological andmorphological characteristics of hybrid tomato designated HMX8148 fruitswhen grown in the same environmental conditions.

In some embodiments, methods of producing said tomato fruit comprise (a)growing the tomato plant from deposited HMX8148 seed to produce a tomatofruit, and (b) harvesting said tomato fruit. In some embodiments, thepresent invention also teaches a tomato fruit produced by the method ofproducing tomato fruit and/or seed as described above. In someembodiments, such fruits have all the physiological and morphologicalcharacteristics of fruits of hybrid tomato designated HMX8148 (e.g.those listed in Table 1) when grown in the same environmentalconditions.

In some embodiments, the present invention teaches methods for producinga tomato seed comprising crossing a first parent tomato plant with asecond parent tomato plant and harvesting the resultant tomato seed,wherein said first parent tomato plant and/or second parent tomato plantis the tomato plant produced from the deposited HMX8148 seed or a tomatoplant having all the characteristics of hybrid tomato HMX8148 as listedin Table 1 including but not limited to as determined at the 5%significance level when grown in the same environmental conditions.

In some embodiments, the present invention teaches methods for producinga tomato seed comprising self-pollinating the tomato plant grown fromthe deposited HMX8148 seed and harvesting the resultant tomato seed.

In some embodiments, the present invention teaches the seed produced byany of the above described methods.

In some embodiments, the present invention teaches methods ofvegetatively propagating the tomato plant grown from the depositedHMX8148 seed, said method comprising collecting a part of a plant grownfrom the deposited HMX8148 seed and regenerating a plant from said part.

In some embodiments, the method further comprises harvesting fruitsand/or seeds from said vegetatively propagated plant. In someembodiments, the method further comprises harvesting a fruit from saidvegetatively propagated plant.

In some embodiments, the present invention teaches the plant and thefruits and/or seeds of plants vegetatively propagated from parts ofplants grown from the deposited HMX8148 seed. In some embodiments, suchplant, fruits and/or seeds have all the physiological and morphologicalcharacteristics of plant, fruits and/or seeds of hybrid tomato HMX8148(e.g. those listed in Table 1) when grown in the same environmentalconditions.

In some embodiments, the present invention teaches methods of producinga tomato plant derived from the hybrid tomato HMX8148. In someembodiment, the methods comprise (a) self-pollinating the plant grownfrom the deposited HMX8148 seed at least once to produce a progeny plantderived from tomato hybrid HMX8148. In some embodiments, the methodfurther comprises (b) crossing the progeny plant derived from tomatohybrid HMX8148 with itself or a second tomato plant to produce a seed ofa progeny plant of a subsequent generation; and; (c) growing the progenyplant of the subsequent generation from the seed, and (d) crossing theprogeny plant of the subsequent generation with itself or a secondtomato plant to produce a tomato plant derived from the hybrid tomatovariety HMX8148. In some embodiments said methods further comprise thestep of: (e) repeating steps (b), (c) and/or (d) for at least 1, 2, 3,4, 5, 6, 7, or more generation to produce a tomato plant derived fromthe hybrid tomato variety HMX8148.

In some embodiments, the present invention teaches methods of producinga tomato plant derived from the hybrid tomato HMX8148, the methodscomprising (a) crossing the plant grown from the deposited HMX8148 seedwith a second tomato plant to produce a progeny plant derived fromhybrid tomato HMX8148. In some embodiments, the method furthercomprises; (b) crossing the progeny plant derived from hybrid tomatoHMX8148 with itself or a second tomato plant to produce a seed of aprogeny plant of a subsequent generation; and; (c) growing the progenyplant of the subsequent generation from the seed; (d) crossing theprogeny plant of the subsequent generation with itself or a secondtomato plant to produce a tomato plant derived from the hybrid tomatovariety HMX8148. In some embodiments said methods further comprise thesteps of: (e) repeating steps (b), (c) and/or (d) for at least 1, 2, 3,4, 5, 6, 7 or more generations to produce a tomato plant derived fromthe hybrid tomato variety HMX8148.

In some embodiments, the present invention teaches plants grown from thedeposited HMX8148 seed wherein said plants comprise a single locusconversion. As used herein, the term “a” or “an” refers to one or moreof that entity; for example, “a single locus conversion” refers to oneor more single locus conversions or at least one single locusconversion. As such, the terms “a” (or “an”), “one or more” and “atleast one” are used interchangeably herein. In addition, reference to“an element” by the indefinite article “a” or “an” does not exclude thepossibility that more than one of the elements are present, unless thecontext clearly requires that there is one and only one of the elements.

In some embodiments, the present invention teaches a method of producinga plant of hybrid tomato designated HMX8148 comprising at least onedesired trait, the method comprising introducing a single locusconversion conferring the desired trait into hybrid tomato designatedHMX8148, whereby a plant of hybrid tomato designated HMX8148 comprisingthe desired trait is produced.

In some embodiments, the present invention teaches a tomato plant,comprising a single locus conversion and essentially all thecharacteristics of hybrid tomato designated HMX8148 listed in Table 1when grown under the same environmental conditions, wherein arepresentative sample of seed of said hybrid has been deposited underNCIMB No. 43896. In other embodiments, the single locus conversion isintroduced into the plant by the use of recurrent selection, mutationbreeding, wherein said mutation breeding selects for a mutation that isspontaneous or artificially induced, backcrossing, pedigree breeding,haploid/double haploid production, marker-assisted selection, genetictransformation, genomic selection, Zinc finger nuclease (ZFN)technology, oligonucleotide directed mutagenesis, cisgenesis,intragenesis, RNA-dependent DNA methylation, agro-infiltration,Transcription Activation-Like Effector Nuclease (TALENs), CRISPR/Cassystem, engineered meganuclease, re-engineered homing endonuclease, andDNA guided genome editing.

In some embodiments, the plant comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,or more single locus conversions. In some embodiments, the plantcomprises no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 single locusconversions, but essentially all the other physiological andmorphological characteristics of hybrid tomato plant HMX8148 listed inTable 1. In some embodiments, the plant comprises at least one singlelocus conversion and essentially all the physiological and morphologicalcharacteristics of hybrid tomato plant HMX8148 listed in Table 1. Inother embodiments, the plant comprises one single locus conversion andessentially all of the other physiological and morphologicalcharacteristics of hybrid tomato plant HMX8148 listed in Table 1.

In some embodiments, said single locus conversion confers said plantswith a trait selected from the group consisting of male sterility, malefertility, herbicide resistance, insect resistance, resistance forbacterial, fungal, mycoplasma or viral disease, enhanced plant qualitysuch as improved drought or salt tolerance, water stress tolerance,improved standability, enhanced plant vigor, improved shelf life,delayed senescence or controlled ripening, increased nutritional qualitysuch as increased sugar content or increased sweetness, increasedtexture, flavor and aroma, improved fruit length and/or size, protectionfor color, fruit shape, uniformity, length or diameter, refinement ordepth lodging resistance, yield and recovery when compared to a suitablecheck/comparison plant. In further embodiments, the single locusconversion confers said plant with herbicide resistance.

In some embodiments, the check plant is a hybrid tomato HMX8148 nothaving said single locus conversion conferring the desired trait(s). Insome embodiments, the at least one single locus conversion is anartificially mutated gene or a gene or nucleotide sequence modifiedthrough the use of New Breeding Techniques.

In some embodiments, the present invention teaches methods of producinga tomato plant, comprising grafting a rootstock or a scion of the hybridtomato plant grown from the deposited HMX8148 seed to another tomatoplant. In some embodiments, the present invention teaches methods forproducing nucleic acids, comprising isolating nucleic acids from theplant grown from the deposited HMX8148 seed, or a part, or a cellthereof. In some embodiments, the present invention teaches methods forproducing a second tomato plant, comprising applying plant breedingtechniques to the plant grown from the deposited HMX8148 seed, or partthereof to produce the second tomato plant.

In some embodiments, the present invention provides a method ofproducing a commodity plant product comprising collecting the commodityplant product from the plant of the present invention. The commodityplant product produced by said method is also part of the presentinvention.

In addition to the exemplary aspects and embodiments described above,further aspects and embodiments will become apparent by study of thefollowing descriptions.

DETAILED DESCRIPTION OF THE INVENTION Definitions

In the description and tables that follow, a number of terms are used.In order to provide a clear and consistent understanding of thespecification and claims, including the scope to be given such terms,the following definitions are provided:

Abscission zone: This is the zone of abscission or area of separation ofthe leaves, flowers, and fruits from the plant. For flower abscission,the resulting zone (or blossom scar) ranges in size, small beingpreferred over large—range small (<10 mm), medium (10-15 mm), large(15-20 mm), very large (>20 mm).

Adaptability: A plant that has adaptability is a plant able to grow wellin different growing conditions (climate, soils, etc.).

Allele: An allele is any of one or more alternative forms of a genewhich relate to one trait or characteristic. In a diploid cell ororganism, the two alleles of a given gene occupy corresponding loci on apair of homologous chromosomes.

Androecious plant: A plant having staminate flowers only.

Backcrossing: Backcrossing is a process in which a breeder repeatedlycrosses hybrid progeny back to one of the parents, for example, afirst-generation hybrid F₁ with one of the parental genotypes of the F₁hybrid.

Blossom scar: This is the remnant scar from the stigmatic surface of theblossom. There is a very broad range in sizes, small is better. Range issmall (<10 mm), medium (10-20 mm), large (20-40 mm) and very large (>40mm).

Cavity: As used herein, cavity refers to the center of the tomato fruitcontaining seeds and maternal tissues. Cavity measurements are made on asingle fruit or recorded as an average of many fruit at harvest maturityand recorded in a convenient unit of measure. Cavity ratings: 1=verypoor (non-marketable), 3=poor (non-marketable), 5=average (marketable)7=very good (much better than industry standards), 9=superior (furtherimprovement not attainable). Cavity evaluations are done based on acombination of the cavity size and the degree of open space in thecavity. Very poor would be open and very large; superior would be verysmall and closed.

Cavity to Diameter ratio: Cavity to Diameter ratio is a measure of thecavity size compared to the overall fruit size of a single fruit or theaverage of many fruit at harvest maturity and recorded in a convenientunit of measure.

Commodity plant product: A “commodity plant product” refers to anycomposition or product that is comprised of material derived from aplant, seed, plant cell, or plant part of the present invention.Commodity plant products may be sold to consumers and can be viable ornonviable. Nonviable commodity products include but are not limited tononviable seeds and grains; processed seeds, seed parts, and plantparts; dehydrated plant tissue, frozen plant tissue, and processed planttissue; seeds and plant parts processed for animal feed for terrestrialand/or aquatic animal consumption, oil, meal, flour, flakes, bran,fiber, paper, tea, coffee, silage, crushed of whole grain, and any otherfood for human or animal consumption; biomasses and fuel products; andraw material in industry.

Collection of seeds: In the context of the present invention acollection of seeds is a grouping of seeds mainly containing similarkind of seeds, for example hybrid seeds having the inbred line of theinvention as a parental line, but that may also contain, mixed togetherwith this first kind of seeds, a second, different kind of seeds, of oneof the inbred parent lines, for example the inbred line of the presentinvention. A commercial bag of hybrid seeds having the inbred line ofthe invention as a parental line and containing also the inbred lineseeds of the invention would be, for example such a collection of seeds.

Determinate tomatoes: Determinate tomatoes are tomato varieties thatcome to fruit all at once, then stop bearing. They are best suited forcommercial growing and mechanical harvesting since they can be harvestedall at once.

Decreased vigor: A plant having a decreased vigor in the presentinvention is a plant that, compared to other plants has a less vigorousappearance for vegetative and/or reproductive characteristics includingshorter plant height, small fruit size, fewer fruit or othercharacteristics.

Earliness: The earliness relates the number of fruits produced from 12to 15 days following the beginning of the harvest: the more fruitsproduced, the more earliness of the plant

Easy to pick fruit: A fruit that is easy to pick is a fruit that easilydetaches from the plant. Once grabbed and twisted, the fruit will breakbetween the peduncle and the stem. For fruits not easy to pick, thepeduncle breaks off the fruits. A fruit that is easy to pick is also afruit that is easily accessible for harvest. When plants have an openplant habit, the fruits are harvested more easily than when the plantshave closed habit.

Enhanced nutritional quality: The nutritional quality of the tomato ofthe present invention can be enhanced by the introduction of severaltraits comprising a higher endosperm sugar content, flesh texture, brix,aroma content and increased sweetness, increased lycopene content of thepeel, etc.

Essentially all the physiological and morphological characteristics: Aplant having essentially all the physiological and morphologicalcharacteristics means a plant having all the physiological andmorphological characteristics of a plant of the present invention,except for desired characteristic(s), which can be indirectly obtainedfrom another plant possessing at least one single locus conversion via aconventional breeding program (such as backcross breeding) or directlyobtained by introduction of at least one single locus conversion via NewBreeding Techniques. In some embodiments, one of the non-limitingexamples for a plant having (and/or comprising) essentially all thephysiological and morphological characteristics shall be a plant havingall the physiological and morphological characteristics of a plant ofthe present invention other than desired, additionaltrait(s)/characteristic(s) conferred by a single locus conversionincluding, but not limited to, a converted or modified gene.

Extended harvest: An extended harvest is a plant that produces fruitsthroughout the harvest season.

Flesh color: In the context of the present invention, the flesh color isthe color of the tomato flesh. Field holding ability: Field holdingability is the ability for fruit quality to maintain even after fruit isripe.

Firm Fruit Exterior. Fruit Firmness subjectively tested under fieldconditions for resistance of fruit exterior against a given pressure.Range is soft, medium, firm and very firm and hard shell.

Grafting: Grafting is the operation by which a rootstock is grafted witha scion. The primary motive for grafting is to avoid damages bysoil-born pest and pathogens when genetic or chemical approaches fordisease management are not available. Grafting a susceptible scion ontoa resistant rootstock can provide a resistant cultivar without the needto breed the resistance into the cultivar. In addition, grafting mayenhance tolerance to abiotic stress, increase yield and result in moreefficient water and nutrient uses.

Good Seed Producer: A plant is a good seed producer when it producesnumerous seeds. For tomato, a good seed producing plant will produce anaverage of 20 grams of seeds during the harvest season.

Gynoecious plant: A plant having pistillate flowers only.

Immunity to disease(s) and or insect(s): A tomato plant which is notsubject to attack or infection by specific disease(s) and or insect(s)is considered immune.

Industrial usage: The industrial usage of the tomato of the presentinvention comprises the use of the tomato fruit for consumption, whetheras fresh products or in canning, freezing or any other industries.

Intermediate resistance to disease(s) and or insect(s): A tomato plantthat restricts the growth and development of specific disease(s) and orinsect(s), but may exhibit a greater range of symptoms or damagecompared to a resistant plant. Intermediate resistant plants willusually show less severe symptoms or damage than susceptible plantvarieties when grown under similar environmental conditions and/orspecific disease(s) and or insect(s) pressure, but may have heavy damageunder heavy pressure. Intermediate resistant tomato plants are notimmune to the disease(s) and or insect(s).

Maturity: In the region of best adaptability, maturity is the number ofdays from transplanting to optimal time for fruit harvest.

Large plant: A large plant has long internodes with a plant height of 75cm and above. It depends on how the plant spreads out horizontally orvertically.

Monecious: The term used to describe a plant variety where each flowerexhibits only one sexual character (either male or female) and eachplant has flowers of both sexes.

New Breeding Techniques: New breeding techniques (NBTs) are said ofvarious new technologies developed and/or used to create newcharacteristics in plants through genetic variation, the aim beingtargeted mutagenesis, targeted introduction of new genes or genesilencing. The following breeding techniques are within the scope ofNBTs: targeted sequence changes facilitated through the use of Zincfinger nuclease (ZFN) technology (ZFN-1, ZFN-2 and ZFN-3, see U.S. Pat.No. 9,145,565, incorporated by reference in its entirety),Oligonucleotide directed mutagenesis (ODM, a.k.a., site-directedmutagenesis), Cisgenesis and intragenesis, epigenetic approaches such asRNA-dependent DNA methylation (RdDM, which does not necessarily changenucleotide sequence but can change the biological activity of thesequence), Grafting (on GM rootstock), Reverse breeding,Agro-infiltration for transient gene expression (agro-infiltration“sensu stricto”, agro-inoculation, floral dip), TranscriptionActivator-Like Effector Nucleases (TALENs, see U.S. Pat. Nos. 8,586,363and 9,181,535, incorporated by reference in their entireties), theCRISPR/Cas system (see U.S. Pat. Nos. 8,697,359; 8,771,945; 8,795,965;8,865,406; 8,871,445; 8,889,356; 8,895,308; 8,906,616; 8,932,814;8,945,839; 8,993,233; and 8,999,641, which are all hereby incorporatedby reference), engineered meganuclease, re-engineered homingendonucleases, DNA guided genome editing (Gao et al., NatureBiotechnology (2016), doi: 10.1038/nbt.3547, incorporated by referencein its entirety), and Synthetic genomics. A major part of today'stargeted genome editing, another designation for New BreedingTechniques, is the applications to induce a DNA double strand break(DSB) at a selected location in the genome where the modification isintended. Directed repair of the DSB allows for targeted genome editing.Such applications can be utilized to generate mutations (e.g., targetedmutations or precise native gene editing) as well as precise insertionof genes (e.g., cisgenes, intragenes, or transgenes). The applicationsleading to mutations are often identified as site-directed nuclease(SDN) technology, such as SDN1, SDN2 and SDN3. For SDN1, the outcome isa targeted, non-specific genetic deletion mutation: the position of theDNA DSB is precisely selected, but the DNA repair by the host cell israndom and results in small nucleotide deletions, additions orsubstitutions. For SDN2, a SDN is used to generate a targeted DSB and aDNA repair template (a short DNA sequence identical to the targeted DSBDNA sequence except for one or a few nucleotide changes) is used torepair the DSB: this results in a targeted and predetermined pointmutation in the desired gene of interest. As to the SDN3, the SDN isused along with a DNA repair template that contains new DNA sequence(e.g. gene). The outcome of the technology would be the integration ofthat DNA sequence into the plant genome. The most likely applicationillustrating the use of SDN3 would be the insertion of cisgenic,intragenic, or transgenic expression cassettes at a selected genomelocation. A complete description of each of these techniques can befound in the report made by the Joint Research Center (JRC) Institutefor Prospective Technological Studies of the European Commission in 2011and titled “New plant breeding techniques—State-of-the-art and prospectsfor commercial development”, which is incorporated by reference in itsentirety.

Number of Boxes per Acre: The Number of Boxes per Acre—6's, 9's, 12's,15's, 18's or 23's refers to the number of fruit that fit into astandard tomato box.

Open Plant Habit: An open plant habit is a plant where the fruits arevisible without moving the leaves. A plant with closed habit will haveits fruit hidden by leaves that have a high density. An average openplant habit will be between the open and closed habit, and the plantwill have medium leaf density. Whether a plant has open habit or closedhabit is based on the whole of the plant. The more erect the plant, themore compact and therefore the closer the habit. In contrast, when theplant is lodging, sprawling on the ground, it leads to a less compactplant, therefore more “open”.

Overall Rating: A final or Overall Rating is assigned to varietyperformance or a varieties characteristic in test or trial situations ofa variety. Overall Rating can range from 1=very poor to 10 excellent.

Oval: Oval is used to describe fruit shape when the length is greaterthan the width and ranges from a slight oval, oval to heavy oval.

Plant adaptability: A plant having good plant adaptability means a plantthat will perform well in different growing conditions and seasons.

Plant cell: As used herein, the term “plant cell” includes plant cellswhether isolated, in tissue culture, or incorporated in a plant or plantpart.

Plant height: Plant height is taken from the top of the soil to the topof the plant leaf canopy and is measured in centimeters or inches.

Plant Part: As used herein, the term “plant part”, “part thereof” or“parts thereof” includes plant cells, plant protoplasts, plant celltissue cultures from which tomato plants can be regenerated, plantcalli, plant clumps and plant cells that are intact in plants or partsof plants, such as embryos, pollens, ovules, flowers, seeds, fruits,rootstocks, scions, stems, roots, anthers, pistils, root tips, leaves,meristematic cells, axillary buds, hypocotyls, cotyledons, ovaries, seedcoats, endosperms and the like. In some embodiments, the plant part atleast comprises at least one cell of said plant. In some embodiments,the plant part is further defined as a pollen, a meristem, a cell or anovule. In some embodiments, a plant regenerated from the plant part hasall of the phenotypic and morphological characteristics of a tomatohybrid of the present invention, including but not limited to asdetermined at the 5% significance level when grown in the sameenvironmental conditions.

Plant Habit: A plant can be an upright plant (also called erect)providing good coverage for the fruit or it can be open with a weakerhabit exposing the fruit

Predicted paste bostwick. The predicted paste bostwick is the calculatednumber with the brix and Bostwick reading using the following formula:Predicted paste bostwick=−11.53+(1.64*juice brix)+(0.5*juice bostwick).

Quantitative Trait Loci (QTL): Quantitative trait loci refer to geneticloci that control to some degree numerically representable traits thatare usually continuously distributed.

Regeneration: Regeneration refers to the development of a plant fromtissue culture.

Resistance to disease(s) and or insect(s): A tomato plant that restrictsthe growth and development of specific disease(s) and or insect(s) undernormal disease(s) and or insect(s) attack pressure when compared tosusceptible plants. These tomato plants can exhibit some symptoms ordamage under heavy disease(s) and or insect(s) pressure. Resistanttomato plants are not immune to the disease(s) and or insect(s).

Rootstock: A rootstock is the lower part of a plant capable of receivinga scion in a grafting process.

RHS: RHS refers to the Royal Horticultural Society of England whichpublishes an official botanical color chart quantitatively identifyingcolors according to a defined numbering system. The chart may bepurchased from Royal Hort. Society Enterprise Ltd. RHS Garden; Wisley,Woking, Surrey GU236QB, UK.

Scion: A scion is the higher part of a plant capable of being graftedonto a rootstock in a grafting process.

Relative maturity or maturity: Maturity is considered the date of theonset of harvest and is classified as Very Early, Early, Mid Early, Mainand Late or specified by recording the date of the onset of harvest. Inthe region of best adaptability, maturity is the number of days fromtransplanting to optimal time for fruit harvest. In this region, amid-early maturity plant is a plant that is harvested approximately 50days after sowing. An early maturity plant would have 45 days fromplanting to harvest while a late maturity plant will have 55 days untilharvest.

Semi-erect habit: A semi-erect plant has a combination of lateral andupright branching and has an intermediate type habit between a prostateplant habit, having laterally growing branching with fruits most of thetime on the ground and an erect plant habit with branching goingstraight up with fruit being off the ground.

Shape: Refers to external fruit shape. Range is Flat Round, Round, roundoval, oval, elongate.

Single locus converted (conversion): Single locus converted (conversion)plants refer to plants which are developed by a plant breeding techniquecalled backcrossing, wherein essentially all the desired morphologicaland physiological characteristics of a plant are recovered in additionto a single locus transferred into the plant via the backcrossingtechnique or via genetic engineering. A single locus converted plant canalso be referred to a plant with a single locus conversion obtainedthough simultaneous and/or artificially induced mutagenesis or throughthe use of New Breeding Techniques described in the present invention.In some embodiments, the single locus converted plant has essentiallyall the desired morphological and physiological characteristics of theoriginal variety in addition to a single locus converted by spontaneousand/or artificially induced mutations, which is introduced and/ortransferred into the plant by the plant breeding techniques such asbackcrossing. In other embodiments, the single locus converted plant hasessentially all the desired morphological and physiologicalcharacteristics of the original variety in addition to a single locus,gene or nucleotide sequence(s) converted, mutated, modified orengineered through the New Breeding Techniques taught herein. In thepresent invention, single locus converted (conversion) can beinterchangeably referred to single gene converted (conversion).

Small plant: A small plant has short internodes with petiole lengths ofapproximately 40 cm and a plant height of 40 to 60 cm. It depends on howthe plant spreads out horizontally or vertically.

Soluble Solids: Soluble solids refer to the percent of solid materialfound in the fruit tissue, the vast majority of which is sugars. Solublesolids are estimated with a refractometer and measured as degrees Brix.Soluble Solids vary with environment. For example, for California summergrowing conditions the following range would apply. Very high (>12.5%),high (11.5-12.5%), medium (10.5-11.5%), low <10.5%).

Susceptible to disease(s) and or insect(s): A tomato plant that issusceptible to disease(s) and or insect(s) is defined as a tomato plantthat has the inability to restrict the growth and development ofspecific disease(s) and or insect(s). Plants that are susceptible willshow damage when infected and are more likely to have heavy damage undermoderate levels of specific disease(s) and or insect(s).

Tolerance to abiotic stresses: A tomato plant that is tolerant toabiotic stresses has the ability to endure abiotic stress withoutserious consequences for growth, appearance and yield.

Uniformity: Uniformity, as used herein, describes the similarity betweenplants or plant characteristics which can be a described by qualitativeor quantitative measurements.

Variety: A plant variety as used by one skilled in the art of plantbreeding means a plant grouping within a single botanical taxon of thelowest known rank which can be defined by the expression of thecharacteristics resulting from a given genotype or combination ofphenotypes, distinguished from any other plant grouping by theexpression of at least one of the said characteristics and considered asa unit with regard to its suitability for being propagated unchanged(International convention for the protection of new varieties ofplants). The term “variety” can be interchangeably used with “cultivar”or “hybrid in the present application”.

Yield (Tomato yield Tons/Acre). The yield in tons/acre is the actualyield of the tomato fruit at harvest.

Tomato Plants

Practically speaking, all cultivated forms of tomato belong to a speciesnow known as Solanum lycopersicum L. This was the originalclassification and is now considered correct and the former designation,Lycopersicon esculentum Miller, is widely used in older literature, butis no longer considered correct. Solanum is a genus within the extremelylarge and diverse family Solanaceae which is considered to consist ofaround 90 genera, including pepper, tobacco and eggplant. The genusLycopersicon has been divide into two subgenera, the esculentum complexwhich contains those species that can easily be crossed with thecommercial tomato and the peruvianum complex which contains thosespecies which are crossed with considerable difficulty (Stevens, M., andRick, C. M. 1986. Genetics and Breeding. In: The Tomato Crop. Ascientific basis for improvement, pp. 35-109. Atherton, J., Rudich, G.(eds.). Chapman and Hall, New York). Due to its value as a crop, L.esculentum Miller has become widely disseminated all over the world.Even if the precise origin of the cultivated tomato is still somewhatunclear, it seems to come from the Americas, being native to Ecuador,Peru and the Galapagos Island and initially cultivated by Aztecs andIncas as early as 700 AD. Mexico appears to have been the site ofdomestication and the source of the earliest introduction.

It is supposed that the cherry tomato, L. esculentum var. cerasiforme,is the direct ancestor of modern cultivated forms.

Tomato is grown for its fruit, widely used as a fresh market orprocessed product. As a crop, tomato is grown commercially whereverenvironmental conditions permit the production of an economically viableyield. In California, the first largest processing tomato market andsecond largest fresh market in the United States, processing tomato areharvested by machine. The majority of fresh market tomatoes areharvested by hand at vine ripe and mature green stage of ripeness. Freshmarket tomatoes are available in the United States year round.Processing tomato season in California is from late June to October.Processing tomato are used in many forms, as canned tomatoes, tomatojuice, tomato sauce, puree, paste or even catsup. Over the 500,000 acresof tomatoes that are grown annually in the US, approximately 40% aregrown for fresh market consumption, the balance are grown forprocessing.

Tomato is a normally simple diploid species with twelve pairs ofdifferentiated chromosomes. However, polyploidy tomato is also part ofthe present invention. The cultivated tomato is self-fertile and almostexclusively self-pollinating. The tomato flowers are hermaphrodites.Commercial cultivars were initially open pollinated. Most have now beenreplaced by better yielding hybrids. Due to its wide dissemination andhigh value, tomato has been intensively bred. This explains why such awide array of tomato is now available. The shape may range from small tolarge, and there are cherry, plum, pear, blocky, round, and beefsteaktypes. Tomatoes may be grouped by the amount of time it takes for theplants to mature fruit for harvest and, in general, the cultivars areconsidered to be early, midseason or late-maturing. Tomatoes can also begrouped by the plant's growth habit; determinate or indeterminate.Determinate plants tend to grow their foliage first, then set flowersthat mature into fruit if pollination is successful. All of the fruitstend to ripen on a plant at about the same time. Indeterminate tomatoesstart out by growing some foliage, then continue to produce foliage andflowers throughout the growing season. These plants will tend to havetomato fruit in different stages of maturity at any given time. Morerecent developments in tomato breeding have led to a wider array offruit color. In addition to the standard red ripe color, tomatoes can becreamy white, lime green, pink, yellow, golden, orange or purple.

Hybrid vigor has been documented in tomatoes and hybrids are gainingmore and more popularity amongst farmers with uniformity of plantcharacteristics.

Hybrid commercial tomato seed is produced by hand pollination. Pollen ofthe male parent is harvested and manually applied to the stigmaticsurface of the female inbred. Prior to and after hand pollination,flowers are covered so that insects do not bring foreign pollen andcreate a mix or impurity. Flowers are tagged to identify pollinatedfruit from which seed will be harvested.

There are numerous steps in the development of any novel, desirableplant germplasm. Plant breeding begins with the analysis and definitionof problems and weaknesses of the current germplasm, the establishmentof program goals, and the definition of specific breeding objectives.The next step is selection of germplasm that possesses the traits tomeet the program goals. The goal is to combine in a single variety orhybrid an improved combination of desirable traits from the parentalgermplasm.

In tomato, these important traits may include increased fruit number,fruit size and fruit weight, higher seed yield, improved color,resistance to diseases and insects, tolerance to drought and heat,better uniformity, higher nutritional value and better agronomicquality, growth rate, high seed germination, seedling vigor, early fruitmaturity, ease of fruit setting, adaptability for soil and climateconditions, firmness, content in soluble solids, acidity and viscosity.With mechanical harvesting of processing tomato, fruit settingconcentration, harvestability and field holding are also very important.

Particularly desirable traits that may be incorporated by this inventionare improved resistance to different viral, fungal, and bacterialpathogens and improved resistance to insect pests. Important diseasesinclude but are not limited to Tomato yellow leaf curl virus, Tomatospot wilt virus, etc. Improved resistance to insect pests is anotherdesirable trait that may be incorporated into new tomato plantsdeveloped by this invention. Insect pests affecting the various speciesof tomato include, but not limited to arthropod pests such as Tutaabsoluta, Frankliniella occidentalis, Bemisia tabaci, etc.

Other desirable traits include traits related to improved tomato fruits.A non-limiting list of fruit phenotypes used during breeding selectioninclude:

-   -   Average of juice bostwick. The juice Bostwick a measurement of        the viscosity. The viscosity or consistency of tomato products        is affected by the degree of concentration of the tomato, the        amount of and extent of degradation of pectin, the size, shape        and quality of the pulp, and probably to a lesser extent, by the        proteins, sugars and other soluble constituents. The viscosity        is measured in Bostwick centimeters by using instruments such as        a Bostwick Consistometer.    -   pH. The pH is a measure of acidity of the fruit puree. A pH        under 4.5 is desirable to prevent bacterial spoilage of finished        products. pH rises as fruit matures.    -   Fruit color. Fruit color is measured as Hunters a/b ratio, where        a represents red/green, positive values are red, negative values        are green and 0 is neutral; b represents yellow/blue, where        positive values are yellow, negative values are blue and 0 is        neutral; a/b represents the intense of redness: large value        represents deep red color, small value represents light or        yellowish red color.    -   Fruit Weight. The weight of a single fruit or the average of        many fruit measured at harvest maturity and recorded in a        convenient unit of measure.    -   Ostwald. The Ostwald is a measurement of serum viscosity whereas        the measurement are taken using an Ostwald viscometer. The serum        is the non-solid portion of a tomato extract after        centrifugation of the tomato puree. The serum viscosity is        affected by the quantity and quality of soluble pectin. Higher        number reflect higher viscosity of the tomato serum.    -   Fruit firmness. The fruit firmness is the resistance to        penetration and is measured using a Digital Durometer Model        DD-4-00 (Rex Gauge Company, Buffalo Grove, Ill., USA). Durometer        readings are taken at 4 locations (each about 90 degrees apart)        on the approximate mid-point of a tomato, with the tomato laying        on its side. From a fruit sample collected at a given location,        the resistance to penetration is measured with the durometer        from 9 individual fruit at 4 locations per fruit (a total of 36        independent measurements). The P5 value is calculated from the        following equation: D-39/10, where D is the value from the        Durometer.        Tomato Breeding

The goal of tomato breeding is to develop new, unique and superiortomato inbred lines and hybrids. The breeder initially selects andcrosses two or more parental lines, followed by repeated selfing andselection, producing many new genetic combinations. Another method usedto develop new, unique and superior tomato inbred lines and hybridsoccurs when the breeder selects and crosses two or more parental linesfollowed by haploid induction and chromosome doubling that result in thedevelopment of dihaploid inbred lines. The breeder can theoreticallygenerate billions of different genetic combinations via crossing,selfing and mutations and the same is true for the utilization of thedihaploid breeding method.

During the development of new tomato inbreds and hybrids, the tomatobreeder uses tomato plants, but also non-commercial tomato plants, suchas plants that may contain characteristics that the breeder has interestin having in its tomato inbreds and hybrids. Such non-commercial tomatoplants could be wild relatives of tomato species.

Each year, the plant breeder selects the germplasm to advance to thenext generation. This germplasm is grown under unique and differentgeographical, climatic and soil conditions, and further selections arethen made, during and at the end of the growing season. The inbred linesdeveloped are unpredictable. This unpredictability is because thebreeder's selection occurs in unique environments, with no control atthe DNA level (using conventional breeding procedures or dihaploidbreeding procedures), and with millions of different possible geneticcombinations being generated. A breeder of ordinary skill in the artcannot predict the final resulting lines the breeder develops, exceptpossibly in a very gross and general fashion. This unpredictabilityresults in the expenditure of large research monies to develop superiornew tomato inbred lines and hybrids.

The development of commercial tomato hybrids requires the development ofhomozygous inbred lines, the crossing of these lines, and the evaluationof the hybrid crosses.

Pedigree breeding and recurrent selection breeding methods are used todevelop inbred lines from breeding populations. Breeding programscombine desirable traits from two or more inbred lines or variousbroad-based sources into breeding pools from which inbred lines aredeveloped by selfing and selection of desired phenotypes or through thedihaploid breeding method followed by the selection of desiredphenotypes. The new inbreds are crossed with other inbred lines and thehybrids from these crosses are evaluated to determine which havecommercial potential.

Choice of breeding or selection methods depends on the mode of plantreproduction, the heritability of the trait(s) being improved, and thetype of cultivar used commercially (e.g., F hybrid cultivar, purelinecultivar, etc.). For highly heritable traits, a choice of superiorindividual plants evaluated at a single location will be effective,whereas for traits with low heritability, selection should be based onmean values obtained from replicated evaluations of families of relatedplants. Popular selection methods commonly include pedigree selection,modified pedigree selection, mass selection, recurrent selection, andbackcross breeding.

i. Pedigree Selection

Pedigree breeding is used commonly for the improvement ofself-pollinating crops or inbred lines of cross-pollinating crops. Twoparents possessing favorable, complementary traits are crossed toproduce an F₁. An F₂ population is produced by selfing one or severalF₁s or by intercrossing two F₁s (sib mating). The dihaploid breedingmethod could also be used. Selection of the best individuals is usuallybegun in the F₂ population; then, beginning in the F₃, the bestindividuals in the best families are selected. Replicated testing offamilies, or hybrid combinations involving individuals of thesefamilies, often follows in the F₄ generation to improve theeffectiveness of selection for traits with low heritability. At anadvanced stage of inbreeding (i.e., F₆ and F₇), the best lines ormixtures of phenotypically similar lines are tested for potential use asparents of new hybrid cultivars. Similarly, the development of newinbred lines through the dihaploid system requires the selection of thebest inbreds followed by two to five years of testing in hybridcombinations in replicated plots.

The single-seed descent procedure in the strict sense refers to plantinga segregating population, harvesting a sample of one seed per plant, andusing the one-seed sample to plant the next generation. When thepopulation has been advanced from the F2 to the desired level ofinbreeding, the plants from which lines are derived will each trace todifferent F2 individuals. The number of plants in a population declineseach generation due to failure of some seeds to germinate or some plantsto produce at least one seed. As a result, not all of the F2 plantsoriginally sampled in the population will be represented by a progenywhen generation advance is completed.

In a multiple-seed procedure, breeders commonly harvest one or morefruit containing seed from each plant in a population and blend themtogether to form a bulk seed lot. Part of the bulked seed is used toplant the next generation and part is put in reserve. The procedure hasbeen referred to as modified single-seed descent or the bulk technique.

The multiple-seed procedure has been used to save labor at harvest. Itis considerably faster than removing one seed from each fruit by handfor the single seed procedure. The multiple-seed procedure also makes itpossible to plant the same number of seeds of a population eachgeneration of inbreeding. Enough seeds are harvested to make up forthose plants that did not germinate or produce seed.

Descriptions of other breeding methods that are commonly used fordifferent traits and crops can be found in one of several referencebooks (e.g., R. W. Allard, 1960, Principles of Plant Breeding, JohnWiley and Son, pp. 115-161; N. W. Simmonds, 1979, Principles of CropImprovement, Longman Group Limited; W. R. Fehr, 1987, Principles of CropDevelopment, Macmillan Publishing Co.; N. F. Jensen, 1988, PlantBreeding Methodology, John Wiley & Sons).

ii. Backcross Breeding

Backcross breeding has been used to transfer genes for a simplyinherited, highly heritable trait into a desirable homozygous cultivaror inbred line which is the recurrent parent. The source of the trait tobe transferred is called the donor parent. The resulting plant isexpected to have the attributes of the recurrent parent (e.g., cultivar)and the desirable trait transferred from the donor parent. After theinitial cross, individuals possessing the phenotype recurrent parent andthe trait of interest from the donor parent are selected and repeatedlycrossed (backcrossed) to the recurrent parent. The resulting plant isexpected to have the attributes of the recurrent parent (e.g., cultivar)and the desirable trait transferred from the donor parent.

When the term hybrid tomato plant is used in the context of the presentinvention, this also includes any hybrid tomato plant where one or moredesired trait has been introduced through backcrossing methods, whethersuch trait is a naturally occurring one, a mutant, a transgenic one or agene or a nucleotide sequence modified by the use of New BreedingTechniques. Backcrossing methods can be used with the present inventionto improve or introduce one or more characteristic into the inbredparental line, thus potentially introducing these traits in to thehybrid tomato plant of the present invention. The term “backcrossing” asused herein refers to the repeated crossing of a hybrid progeny back tothe recurrent parent, i.e., backcrossing one, two, three, four, five,six, seven, eight, nine, or more times to the recurrent parent. Theparental tomato plant which contributes the gene or the genes for thedesired characteristic is termed the nonrecurrent or donor parent. Thisterminology refers to the fact that the nonrecurrent parent is used onetime in the backcross protocol and therefore does not recur. Theparental tomato plant to which the gene or genes from the nonrecurrentparent are transferred is known as the recurrent parent as it is usedfor several rounds in the backcrossing protocol.

In a typical backcross protocol, the original inbred of interest(recurrent parent) is crossed to or by a second inbred (nonrecurrentparent) that carries the gene or genes of interest to be transferred.The resulting progeny from this cross are then crossed again to or bythe recurrent parent and the process is repeated until a tomato plant isobtained wherein all the desired morphological and physiologicalcharacteristics of the recurrent parent are recovered in the convertedplant, generally determined at a 5% significance level when grown in thesame environmental conditions, in addition to the gene or genestransferred from the nonrecurrent parent. It has to be noted that some,one, two, three or more, self-pollination and growing of populationmight be included between two successive backcrosses. Indeed, anappropriate selection in the population produced by theself-pollination, i.e. selection for the desired trait and physiologicaland morphological characteristics of the recurrent parent might beequivalent to one, two or even three additional backcrosses in acontinuous series without rigorous selection, saving then time, moneyand effort to the breeder. A non-limiting example of such a protocolwould be the following: a) the first generation F1 produced by the crossof the recurrent parent A by the donor parent B is backcrossed to parentA, b) selection is practiced for the plants having the desired trait ofparent B, c) selected plant are self-pollinated to produce a populationof plants where selection is practiced for the plants having the desiredtrait of parent B and physiological and morphological characteristics ofparent A, d) the selected plants are backcrossed one, two, three, four,five, six, seven, eight, nine, or more times to parent A to produceselected backcross progeny plants comprising the desired trait of parentB and the physiological and morphological characteristics of parent A.Step (c) may or may not be repeated and included between the backcrossesof step (d).

The selection of a suitable recurrent parent is an important step for asuccessful backcrossing procedure. The goal of a backcross protocol isto alter or substitute one or more trait(s) or characteristic(s) in theoriginal inbred parental line in order to find it then in the hybridmade thereof. To accomplish this, a gene or genes of the recurrentinbred is modified or substituted with the desired gene or genes fromthe nonrecurrent parent, while retaining essentially all the rest of thedesired genetic, and therefore the desired physiological andmorphological, constitution of the original inbred. The choice of theparticular nonrecurrent parent will depend on the purpose of thebackcross; one of the major purposes is to add some commerciallydesirable, agronomically important trait(s) to the plant. The exactbackcrossing protocol will depend on the characteristic(s) or trait(s)being altered to determine an appropriate testing protocol. Althoughbackcrossing methods are simplified when the characteristic beingtransferred is a single gene and dominant allele, multiple genes andrecessive allele(s) may also be transferred and therefore, backcrossbreeding is by no means restricted to character(s) governed by one or afew genes. In fact, the number of genes might be less important that theidentification of the character(s) in the segregating population. Inthis instance it may then be necessary to introduce a test of theprogeny to determine if the desired characteristic(s) has beensuccessfully transferred. Such tests encompass visual inspection, simplecrossing, but also follow up of the characteristic(s) throughgenetically associated markers and molecular assisted breeding tools.For example, selection of progeny containing the transferred trait isdone by direct selection, visual inspection for a trait associated witha dominant allele, while the selection of progeny for a trait that istransferred via a recessive allele, such as the orange fruit colorcharacteristic in tomato, requires selfing the progeny or usingmolecular markers to determine which plant carry the recessiveallele(s).

Many single gene traits have been identified that are not regularlyselected for in the development of a new parental inbred of a hybridtomato plant according to the invention but that can be improved bybackcrossing techniques. Single gene traits may or may not betransgenic. Examples of these traits include but are not limited to,male sterility (such as the ms1, ms2, ms3, ms4 or ms5 genes), herbicideresistance (such as bar or PAT genes), gynoecia (such as the g gene),resistance for bacterial, fungal (genes Cf for resistance toCladosporium fulvum) or viral disease (gene Ty for resistance to TomatoYellow Leaf Curl Virus (TYLCV), genes Tm-1, Tm-2 and Tm2² for theresistance to the tomato mosaic tobamovirus (ToMV)), insect resistance(gene Mi for resistance to nematodes), increased brix by introduction ofspecific alleles such as the hir4 allele from Lycopersicon hirsutum,high lycopene by using the dg mutant as described in U.S. Ser. No.10/587,789, improved shelf life by using mutants such as the rin(ripening inhibitor), nor (non-ripening) or cnr (colorless non ripening)alleles, increased firmness or slower softening of the fruits due, forexample in a mutation in an expansin gene, absence of gel (i.e. fruitshaving a cavity area which is solid and lacks a gel or liquid contentmale) by the use of the PSAF allele, fertility, enhanced nutritionalquality, enhanced sugar content, yield stability and yield enhancement.These genes are generally inherited through the nucleus. Some knownexceptions to this are the genes for male sterility, some of which areinherited cytoplasmically, but still act as single gene traits. Severalof these single gene traits are described in U.S. Pat. Nos. 5,777,196;5,948,957 and 5,969,212, the disclosures of which are specificallyhereby incorporated by reference.

In 1981, the backcross method of breeding counted for 17% of the totalbreeding effort for inbred line development in the United States,accordingly to, Hallauer, A. R. et al. (1988) “Corn Breeding” Corn andCorn Improvement, No. 18, pp. 463-481.

The backcross breeding method provides a precise way of improvingvarieties that excel in a large number of attributes but are deficientin a few characteristics. (Page 150 of the Pr. R. W. Allard's 1960 book,published by John Wiley & Sons, Inc., Principles of Plant Breeding). Themethod makes use of a series of backcrosses to the variety to beimproved during which the character or the characters in whichimprovement is sought is maintained by selection. At the end of thebackcrossing the gene or genes being transferred unlike all other genes,will be heterozygous. Selfing after the last backcross produceshomozygosity for this gene pair(s) and, coupled with selection, willresult in a parental line of a hybrid variety with exactly oressentially the same adaptation, yielding ability and qualitycharacteristics of the recurrent parent but superior to that parent inthe particular characteristic(s) for which the improvement program wasundertaken. Therefore, this method provides the plant breeder with ahigh degree of genetic control of this work.

The method is scientifically exact because the morphological andagricultural features of the improved variety could be described inadvance and because a similar variety could, if it were desired, be breda second time by retracing the same steps (Briggs, “Breeding wheatsresistant to bunt by the backcross method”, 1930 Jour. Amer. Soc.Agron., 22: 289-244).

Backcrossing is a powerful mechanism for achieving homozygosity and anypopulation obtained by backcrossing must rapidly converge on thegenotype of the recurrent parent. When backcrossing is made the basis ofa plant breeding program, the genotype of the recurrent parent will betheoretically modified only with regards to genes being transferred,which are maintained in the population by selection.

Successful backcrosses are, for example, the transfer of stem rustresistance from ‘Hope’ wheat to ‘Bart wheat’ and even pursuing thebackcrosses with the transfer of bunt resistance to create ‘Bart 38’,having both resistances. Also highlighted by Allard is the successfultransfer of mildew, leaf spot and wilt resistances in California Commonalfalfa to create ‘Caliverde’. This new ‘Caliverde’ variety producedthrough the backcross process is indistinguishable from CaliforniaCommon except for its resistance to the three named diseases.

One of the advantages of the backcross method is that the breedingprogram can be carried out in almost every environment that will allowthe development of the character being transferred or when usingmolecular markers that can identify the trait of interest.

The backcross technique is not only desirable when breeding for diseaseresistance but also for the adjustment of morphological characters,color characteristics and simply inherited quantitative characters suchas earliness, plant height and seed size and shape. In this regard, amedium grain type variety, ‘Calady’, has been produced by Jones andDavis. As dealing with quantitative characteristics, they selected thedonor parent with the view of sacrificing some of the intensity of thecharacter for which it was chosen, i.e. grain size. ‘Lady Wright’, along grain variety was used as the donor parent and ‘Coloro’, a shortgrain one as the recurrent parent. After four backcrosses, the mediumgrain type variety ‘Calady’ was produced.

iii. Open-Pollinated Populations

The improvement of open-pollinated populations of such crops as rye,many maizes and sugar beets, herbage grasses, legumes such as alfalfaand clover, and tropical tree crops such as cacao, coconuts, oil palmand some rubber, depends essentially upon changing gene-frequenciestowards fixation of favorable alleles while maintaining a high (but farfrom maximal) degree of heterozygosity.

Uniformity in such populations is impossible and trueness-to-type in anopen-pollinated variety is a statistical feature of the population as awhole, not a characteristic of individual plants. Thus, theheterogeneity of open-pollinated populations contrasts with thehomogeneity (or virtually so) of inbred lines, clones and hybrids.

Population improvement methods fall naturally into two groups, thosebased on purely phenotypic selection, normally called mass selection,and those based on selection with progeny testing. Interpopulationimprovement utilizes the concept of open breeding populations; allowinggenes to flow from one population to another. Plants in one population(cultivar, strain, ecotype, or any germplasm source) are crossed eithernaturally (e.g., by wind) or by hand or by bees (commonly Apis melliferaL. or Megachile rotundata F.) with plants from other populations.Selection is applied to improve one (or sometimes both) population(s) byisolating plants with desirable traits from both sources.

There are basically two primary methods of open-pollinated populationimprovement.

First, there is the situation in which a population is changed en masseby a chosen selection procedure. The outcome is an improved populationthat is indefinitely propagated by random-mating within itself inisolation.

Second, the synthetic variety attains the same end result as populationimprovement, but is not itself propagated as such; it has to bereconstructed from parental lines or clones. These plant breedingprocedures for improving open-pollinated populations are well known tothose skilled in the art and comprehensive reviews of breedingprocedures routinely used for improving cross-pollinated plants areprovided in numerous texts and articles, including: Allard, Principlesof Plant Breeding, John Wiley & Sons, Inc. (1960); Simmonds, Principlesof Crop Improvement, Longman Group Limited (1979); Hall auer andMiranda, Quantitative Genetics in Maize Breeding, Iowa State UniversityPress (1981); and, Jensen, Plant Breeding Methodology, John Wiley &Sons, Inc. (1988).

A) Mass Selection

Mass and recurrent selections can be used to improve populations ofeither self- or cross-pollinating crops. A genetically variablepopulation of heterozygous individuals is either identified or createdby intercrossing several different parents. The best plants are selectedbased on individual superiority, outstanding progeny, or excellentcombining ability. The selected plants are intercrossed to produce a newpopulation in which further cycles of selection are continued. In massselection, desirable individual plants are chosen, harvested, and theseed composited without progeny testing to produce the followinggeneration. Since selection is based on the maternal parent only, andthere is no control over pollination, mass selection amounts to a formof random mating with selection. As stated above, the purpose of massselection is to increase the proportion of superior genotypes in thepopulation.

B) Synthetics

A synthetic variety is produced by intercrossing a number of genotypesselected for good combining ability in all possible hybrid combinations,with subsequent maintenance of the variety by open pollination. Whetherparents are (more or less inbred) seed-propagated lines, as in somesugar beet and beans (Vicia) or clones, as in herbage grasses, cloversand alfalfa, makes no difference in principle. Parents are selected ongeneral combining ability, sometimes by test crosses or toperosses, moregenerally by polycrosses. Parental seed lines may be deliberately inbred(e.g. by selfing or sib crossing). However, even if the parents are notdeliberately inbred, selection within lines during line maintenance willensure that some inbreeding occurs. Clonal parents will, of course,remain unchanged and highly heterozygous.

Whether a synthetic can go straight from the parental seed productionplot to the farmer or must first undergo one or more cycles ofmultiplication depends on seed production and the scale of demand forseed. In practice, grasses and clovers are generally multiplied once ortwice and are thus considerably removed from the original synthetic.

While mass selection is sometimes used, progeny testing is generallypreferred for polycrosses, because of their operational simplicity andobvious relevance to the objective, namely exploitation of generalcombining ability in a synthetic.

The number of parental lines or clones that enters a synthetic varieswidely. In practice, numbers of parental lines range from 10 to severalhundred, with 100-200 being the average. Broad based synthetics formedfrom 100 or more clones would be expected to be more stable during seedmultiplication than narrow based synthetics.

iv. Hybrids

A hybrid is an individual plant resulting from a cross between parentsof differing genotypes. Commercial hybrids are now used extensively inmany crops, including corn (maize), sorghum, sugarbeet, sunflower,broccoli and tomato as well as leafy vegetables such as lettuce. Hybridscan be formed in a number of different ways, including by crossing twoparents directly (single cross hybrids), by crossing a single crosshybrid with another parent (three-way or triple cross hybrids), or bycrossing two different hybrids (four-way or double cross hybrids).

Strictly speaking, most individuals in an out breeding (i.e.,open-pollinated) population are hybrids, but the term is usuallyreserved for cases in which the parents are individuals whose genomesare sufficiently distinct for them to be recognized as different speciesor subspecies. Hybrids may be fertile or sterile depending onqualitative and/or quantitative differences in the genomes of the twoparents. Heterosis, or hybrid vigor, is usually associated withincreased heterozygosity that results in increased vigor of growth,survival, and fertility of hybrids as compared with the parental linesthat were used to form the hybrid. Maximum heterosis is usually achievedby crossing two genetically different, highly inbred lines.

Hybrid commercial tomato seed can be produced by controlled handpollination. The male flowers from the male plants are harvested andused to pollinate the stigmatic surface of the female flowers on thefemale plants. Prior to, and after hand pollination, flowers are coveredso that insects do not bring foreign pollen and create a mix orimpurity. Flowers are tagged to identify pollinated fruit from whichseed will be harvested.

Once the inbreds that give the best hybrid performance have beenidentified, the hybrid seed can be reproduced indefinitely as long asthe homogeneity of the inbred parent is maintained. A single-crosshybrid is produced when two inbred lines are crossed to produce the F1progeny. A double-cross hybrid is produced from four inbred linescrossed in pairs (A×B and C×D) and then the two F1 hybrids are crossedagain (A×B)×(C×D). Much of the hybrid vigor and uniformity exhibited byF1 hybrids is lost in the next generation (F2). Consequently, seed fromF2 hybrid varieties is not used for planting stock.

The production of hybrids is a well-developed industry, involving theisolated production of both the parental lines and the hybrids whichresult from crossing those lines. For a detailed discussion of thehybrid production process, see, e.g., Wright, Commercial Hybrid SeedProduction 8:161-176, In Hybridization of Crop Plants.

v. Bulk Segregation Analysis (BSA)

BSA, a.k.a. bulked segregation analysis, or bulk segregant analysis, isa method described by Michelmore et al. (Michelmore et al., 1991,Identification of markers linked to disease-resistance genes by bulkedsegregant analysis: a rapid method to detect markers in specific genomicregions by using segregating populations. Proceedings of the NationalAcademy of Sciences, USA, 99:9828-9832) and Quarrie et al. (Quarrie etal., 1999, Journal of Experimental Botany, 50(337):1299-1306).

For BSA of a trait of interest, parental lines with certain differentphenotypes are chosen and crossed to generate F2, doubled haploid orrecombinant inbred populations with QTL analysis. The population is thenphenotyped to identify individual plants or lines having high or lowexpression of the trait. Two DNA bulks are prepared, one from theindividuals having one phenotype (e.g., resistant to virus), and theother from the individuals having reversed phenotype (e.g., susceptibleto virus), and analyzed for allele frequency with molecular markers.Only a few individuals are required in each bulk (e.g., 10 plants each)if the markers are dominant (e.g., RAPDs). More individuals are neededwhen markers are co-dominant (e.g., RFLPs, SNPs or SSRs). Markers linkedto the phenotype can be identified and used for breeding or QTL mapping.

vi. Hand-Pollination Method

Hand pollination describes the crossing of plants via the deliberatefertilization of female ovules with pollen from a desired male parentplant. In some embodiments the donor or recipient female parent and thedonor or recipient male parent line are planted in the same field. Theinbred male parent can be planted earlier than the female parent toensure adequate pollen supply at the pollination time. In someembodiments, the male parent and female parent can be planted at a ratioof 1 male parent to 4-10 female parents. The male parent may be plantedat the top of the field for efficient male flower collection duringpollination. Pollination is started when the female parent flower isready to be fertilized. Female flower buds that are ready to open in thefollowing days are identified, covered with paper cups or small paperbags that prevent bee or any other insect from visiting the femaleflowers, and marked with any kind of material that can be easily seenthe next morning. In some embodiments, this process is best done in theafternoon. The male flowers of the male parent are collected in theearly morning before they are open and visited by pollinating insects.The covered female flowers of the female parent, which have opened, areun-covered and pollinated with the collected fresh male flowers of themale parent, starting as soon as the male flower sheds pollen. Thepollinated female flowers are again covered after pollination to preventbees and any other insects visit. The pollinated female flowers are alsomarked. The marked fruits are harvested. In some embodiments, the malepollen used for fertilization has been previously collected and stored.

vii. Bee-Pollination Method

Using the bee-pollination method, the parent plants are usually plantedwithin close proximity. In some embodiments more female plants areplanted to allow for a greater production of seed. Breeding of dioeciousspecies can also be done by growing equal amount of each parent plant.Insects are placed in the field or greenhouses for transfer of pollenfrom the male parent to the female flowers of the female parent. In someembodiments, fruits set after the introduction of the beehives can bemarked for later collection.

viii. Targeting Induced Local Lesions in Genomes (TILLING)

Breeding schemes of the present application can include crosses withTILLING® plant lines. TILLING® is a method in molecular biology thatallows directed identification of mutations in a specific gene. TILLING®was introduced in 2000, using the model plant Arabidopsis thaliana.TILLING® has since been used as a reverse genetics method in otherorganisms such as zebrafish, corn, wheat, rice, soybean, tomato andlettuce.

The method combines a standard and efficient technique of mutagenesiswith a chemical mutagen (e.g., Ethyl methanesulfonate (EMS)) with asensitive DNA screening-technique that identifies single base mutations(also called point mutations) in a target gene. EcoTILLING is a methodthat uses TILLING® techniques to look for natural mutations inindividuals, usually for population genetics analysis (see Comai, etal., 2003 The Plant Journal 37, 778-786; Gilchrist et al. 2006 Mol.Ecol. 15, 1367-1378; Mejlhede et al. 2006 Plant Breeding 125, 461-467;Nieto et al. 2007 BMC Plant Biology 7, 34-42, each of which isincorporated by reference hereby for all purposes). DEcoTILLING is amodification of TILLING® and EcoTILLING which uses an inexpensive methodto identify fragments (Garvin et al., 2007, DEco-TILLING: An inexpensivemethod for SNP discovery that reduces ascertainment bias. MolecularEcology Notes 7, 735-746).

The TILLING® method relies on the formation of heteroduplexes that areformed when multiple alleles (which could be from a heterozygote or apool of multiple homozygotes and heterozygotes) are amplified in a PCR,heated, and then slowly cooled. As DNA bases are not pairing at themismatch of the two DNA strands (the induced mutation in TILLING® or thenatural mutation or SNP in EcoTILLING), they provoke a shape change inthe double strand DNA fragment which is then cleaved by single strandednucleases. The products are then separated by size on several differentplatforms.

Several TILLING® centers exists over the world that focus onagriculturally important species: UC Davis (USA), focusing on Rice;Purdue University (USA), focusing on Maize; University of BritishColumbia (CA), focusing on Brassica napus; John Innes Centre (UK),focusing on Brassica raga; Fred Hutchinson Cancer Research, focusing onArabidopsis; Southern Illinois University (USA), focusing on Soybean;John Innes Centre (UK), focusing on Lotus and Medicago; and INRA(France), focusing on Pea and Tomato.

More detailed description on methods and compositions on TILLING® can befound in U.S. Pat. No. 5,994,075, US 2004/0053236 A1, WO 2005/055704,and WO 2005/048692, each of which is hereby incorporated by referencefor all purposes.

Thus, in some embodiments, the breeding methods of the presentdisclosure include breeding with one or more TILLING plant lines withone or more identified mutations.

ix. Mutation Breeding

Mutation breeding is another method of introducing new variation andsubsequent traits into tomato plants. Mutations that occur spontaneouslyor are artificially induced can be useful sources of variability for aplant breeder. The goal of artificial mutagenesis is to increase therate of mutation for a desired characteristic. Mutation rates can beincreased by many different means or mutating agents includingtemperature, long-term seed storage, tissue culture conditions,radiation (such as X-rays, Gamma rays, neutrons, Beta radiation, orultraviolet radiation), chemical mutagens (such as base analogs like5-bromo-uracil), antibiotics, alkylating agents (such as sulfurmustards, nitrogen mustards, epoxides, ethyleneamines, sulfates,sulfonates, sulfones, or lactones), azide, hydroxylamine, nitrous acidor acridines. Once a desired trait is observed through mutagenesis thetrait may then be incorporated into existing germplasm by traditionalbreeding techniques. Details of mutation breeding can be found in W. R.Fehr, 1993, Principles of Cultivar Development, Macmillan Publishing Co.

New breeding techniques such as the ones involving the uses ofengineered nuclease to enhance the efficacy and precision of geneediting in combination with oligonucleotides including, but not limitedto Zinc Finger Nucleases (ZFN), TAL effector nucleases (TALENs) andclustered regularly interspaced short palindromic repeats(CRISPR)-associated endonuclease Cas9 (CRISPR-Cas9) shall also be usedto generate genetic variability and introduce new traits into tomatovarieties.

x. Double Haploids and Chromosome Doubling

One way to obtain homozygous plants without the need to cross twoparental lines followed by a long selection of the segregating progeny,and/or multiple backcrossing is to produce haploids and then double thechromosomes to form doubled haploids. Haploid plants can occurspontaneously, or may be artificially induced via chemical treatments orby crossing plants with inducer lines (Seymour et al. 2012, PNAS vol.109, pg. 4227-4232; Zhang et al., 2008 Plant Cell Rep. December 27(12)1851-60). The production of haploid progeny can occur via a variety ofmechanisms which can affect the distribution of chromosomes duringgamete formation. The chromosome complements of haploids sometimesdouble spontaneously to produce homozygous doubled haploids (DHs).Mixoploids, which are plants which contain cells having differentploidies, can sometimes arise and may represent plants that areundergoing chromosome doubling so as to spontaneously produce doubledhaploid tissues, organs, shoots, floral parts or plants. Another commontechnique is to induce the formation of double haploid plants with achromosome doubling treatment such as colchicine (El-Hennawy et al.,2011 Vol 56, issue 2 pg. 63-72; Doubled Haploid Production in CropPlants 2003 edited by Maluszynski ISBN 1-4020-1544-5). The production ofdoubled haploid plants yields highly uniform inbred lines and isespecially desirable as an alternative to sexual inbreeding oflonger-generation crops. By producing doubled haploid progeny, thenumber of possible gene combinations for inherited traits is moremanageable. Thus, an efficient doubled haploid technology cansignificantly reduce the time and the cost of inbred and cultivardevelopment.

xi. Protoplast Fusion

In another method for breeding plants, protoplast fusion can also beused for the transfer of trait-conferring genomic material from a donorplant to a recipient plant. Protoplast fusion is an induced orspontaneous union, such as a somatic hybridization, between two or moreprotoplasts (cells of which the cell walls are removed by enzymatictreatment) to produce a single bi- or multi-nucleate cell. The fusedcell that may even be obtained with plant species that cannot beinterbred in nature is tissue cultured into a hybrid plant exhibitingthe desirable combination of traits.

xii. Embryo Rescue

Alternatively, embryo rescue may be employed in the transfer ofresistance-conferring genomic material from a donor plant to a recipientplant. Embryo rescue can be used as a procedure to isolate embryos fromcrosses to rapidly move to the next generation of backcrossing orselfing or wherein plants fail to produce viable seed. In this process,the fertilized ovary or immature seed of a plant is tissue cultured tocreate new plants (see Pierik, 1999, In Vitro Culture of Higher Plants,Springer, ISBN 079235267X, 978-0792352679, which is incorporated hereinby reference in its entirety).

Grafting

Grafting is a process that has been used for many years in crops such ascucurbitacea, but only more recently for some commercial watermelon andtomato production. Grafting may be used to provide a certain level ofresistance to telluric pathogens such as Phytophthora or to certainnematodes. Grafting is therefore intended to prevent contact between theplant or variety to be cultivated and the infested soil. The variety ofinterest used as the graft or scion, optionally an F1 hybrid, is graftedonto the resistant plant used as the rootstock. The resistant rootstockremains healthy and provides, from the soils, the normal supply for thegraft that it isolates from the diseases. In some recent developments,it has also been shown that some rootstocks are also able to improve theagronomic value for the grafted plant and in particular the equilibriumbetween the vegetative and generative development that are alwaysdifficult to balance in tomato cultivation.

Breeding Evaluation

Each breeding program can include a periodic, objective evaluation ofthe efficiency of the breeding procedure. Evaluation criteria varydepending on the goal and objectives, but should include gain fromselection per year based on comparisons to an appropriate standard,overall value of the advanced breeding lines, and number of successfulcultivars produced per unit of input (e.g., per year, per dollarexpended, etc.).

Promising advanced breeding lines are thoroughly tested per se and inhybrid combination and compared to appropriate standards in environmentsrepresentative of the commercial target area(s). The best lines arecandidates for use as parents in new commercial cultivars; those stilldeficient in a few traits may be used as parents to produce newpopulations for further selection or in a backcross program to improvethe parent lines for a specific trait.

In some embodiments, the plants are selected on the basis of one or morephenotypic traits. Skilled persons will readily appreciate that suchtraits include any observable characteristic of the plant, including forexample growth rate, vigor, plant health, maturity, branching, plantheight, leaf coverage, weight, total yield, color, taste, sugar levels,aroma, changes in the production of one or more compounds by the plant(including for example, metabolites, proteins, drugs, carbohydrates,oils, and any other compounds).

A most difficult task is the identification of individuals that aregenetically superior, because for most traits the true genotypic valueis masked by other confounding plant traits or environmental factors.One method of identifying a superior plant is to observe its performancerelative to other experimental plants and to a widely grown standardcultivar. If a single observation is inconclusive, replicatedobservations provide a better estimate of its genetic worth.

Proper testing should detect any major faults and establish the level ofsuperiority or improvement over current cultivars. In addition toshowing superior performance, there must be a demand for a new cultivarthat is compatible with industry standards or which creates a newmarket. The introduction of a new cultivar will incur additional coststo the seed producer, the grower, processor and consumer for specialadvertising and marketing, altered seed and commercial productionpractices, and new product utilization. The testing preceding release ofa new cultivar should take into consideration research and developmentcosts as well as technical superiority of the final cultivar. Forseed-propagated cultivars, it must be feasible to produce seed easilyand economically.

It should be appreciated that in certain embodiments, plants may beselected based on the absence, suppression or inhibition of a certainfeature or trait (such as an undesirable feature or trait) as opposed tothe presence of a certain feature or trait (such as a desirable featureor trait).

Selecting plants based on genotypic information is also envisaged (forexample, including the pattern of plant gene expression, genotype, orpresence of genetic markers). Where the presence of one or more geneticmarker is assessed, the one or more marker may already be known and/orassociated with a particular characteristic of a plant; for example, amarker or markers may be associated with an increased growth rate ormetabolite profile. This information could be used in combination withassessment based on other characteristics in a method of the disclosureto select for a combination of different plant characteristics that maybe desirable. Such techniques may be used to identify novel quantitativetrait loci (QTLs). By way of example, plants may be selected based ongrowth rate, size (including but not limited to weight, height, leafsize, stem size, branching pattern, or the size of any part of theplant), general health, survival, tolerance to adverse physicalenvironments and/or any other characteristic, as described hereinbefore.

Further non-limiting examples include selecting plants based on: speedof seed germination; quantity of biomass produced; increased root,and/or leaf/shoot growth that leads to an increased yield (fruit) orbiomass production; effects on plant growth that results in an increasedseed yield for a crop; effects on plant growth which result in anincreased yield; effects on plant growth that lead to an increasedresistance or tolerance to disease including fungal, viral or bacterialdiseases, to mycoplasma, or to pests such as insects, mites or nematodesin which damage is measured by decreased foliar symptoms such as theincidence of bacterial or fungal lesions, or area of damaged foliage orreduction in the numbers of nematode cysts or galls on plant roots, orimprovements in plant yield in the presence of such plant pests anddiseases; effects on plant growth that lead to increased metaboliteyields; effects on plant growth that lead to improved aesthetic appealwhich may be particularly important in plants grown for their form,color or taste, for example the color intensity of tomato exocarp (skin)of said fruit.

Molecular Breeding Evaluation Techniques

Selection of plants based on phenotypic or genotypic information may beperformed using techniques such as, but not limited to: high through-putscreening of chemical components of plant origin, sequencing techniquesincluding high through-put sequencing of genetic material, differentialdisplay techniques (including DDRT-PCR, and DD-PCR), nucleic acidmicroarray techniques, RNA-seq (Transcriptome Sequencing), qRTPCR(quantitative real time PCR).

In one embodiment, the evaluating step of a plant breeding programinvolves the identification of desirable traits in progeny plants.Progeny plants can be grown in, or exposed to conditions designed toemphasize a particular trait (e.g. drought conditions for droughttolerance, lower temperatures for freezing tolerant traits). Progenyplants with the highest scores for a particular trait may be used forsubsequent breeding steps.

In some embodiments, plants selected from the evaluation step canexhibit a 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%,65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 120% or more improvement in aparticular plant trait compared to a control plant.

In other embodiments, the evaluating step of plant breeding comprisesone or more molecular biological tests for genes or other markers. Forexample, the molecular biological test can involve probe hybridizationand/or amplification of nucleic acid (e.g., measuring nucleic aciddensity by Northern or Southern hybridization, PCR) and/or immunologicaldetection (e.g., measuring protein density, such as precipitation andagglutination tests, ELISA (e.g., Lateral Flow test or DAS-ELISA),Western blot, immune labeling, immunosorbent electron microscopy (ISEM),and/or dot blot).

The procedure to perform a nucleic acid hybridization, an amplificationof nucleic acid (e.g., PCR, RT-PCR) or an immunological detection (e.g.,precipitation and agglutination tests, ELISA (e.g., Lateral Flow test orDAS-ELISA), Western blot, RIA, immunogold or immunofluorescent labeling,immunosorbent electron microscopy (ISEM), and/or dot blot tests) areperformed as described elsewhere herein and well-known by one skilled inthe art.

In one embodiment, the evaluating step comprises PCR (semi-quantitativeor quantitative), wherein primers are used to amplify one or morenucleic acid sequences of a desirable gene, or a nucleic acid associatedwith said gene, or QTL or a desirable trait (e.g., a co-segregatingnucleic acid, or other marker).

In another embodiment, the evaluating step comprises immunologicaldetection (e.g., precipitation and agglutination tests, ELISA (e.g.,Lateral Flow test or DAS-ELISA), Western blot, MA, immuno labeling(gold, fluorescent, or other detectable marker), immunosorbent electronmicroscopy (ISEM), and/or dot blot), wherein one or more gene ormarker-specific antibodies are used to detect one or more desirableproteins. In one embodiment, said specific antibody is selected from thegroup consisting of polyclonal antibodies, monoclonal antibodies,antibody fragments, and combination thereof.

Reverse Transcription Polymerase Chain Reaction (RT-PCR) can be utilizedin the present disclosure to determine expression of a gene to assistduring the selection step of a breeding scheme. It is a variant ofpolymerase chain reaction (PCR), a laboratory technique commonly used inmolecular biology to generate many copies of a DNA sequence, a processtermed “amplification”. In RT-PCR, however, RNA strand is first reversetranscribed into its DNA complement (complementary DNA, or cDNA) usingthe enzyme reverse transcriptase, and the resulting cDNA is amplifiedusing traditional or real-time PCR.

RT-PCR utilizes a pair of primers, which are complementary to a definedsequence on each of the two strands of the mRNA. These primers are thenextended by a DNA polymerase and a copy of the strand is made after eachcycle, leading to logarithmic amplification.

RT-PCR includes three major steps. The first step is the reversetranscription (RT) where RNA is reverse transcribed to cDNA using areverse transcriptase and primers. This step is very important in orderto allow the performance of PCR since DNA polymerase can act only on DNAtemplates. The RT step can be performed either in the same tube with PCR(one-step PCR) or in a separate one (two-step PCR) using a temperaturebetween 40° C. and 60° C., depending on the properties of the reversetranscriptase used.

The next step involves the denaturation of the dsDNA at 95° C., so thatthe two strands separate and the primers can bind again at lowertemperatures and begin a new chain reaction. Then, the temperature isdecreased until it reaches the annealing temperature which can varydepending on the set of primers used, their concentration, the probe andits concentration (if used), and the cation concentration. The mainconsideration, of course, when choosing the optimal annealingtemperature is the melting temperature (Tm) of the primers and probes(if used). The annealing temperature chosen for a PCR depends directlyon length and composition of the primers. This is the result of thedifference of hydrogen bonds between A-T (2 bonds) and G-C (3 bonds). Anannealing temperature about 5 degrees below the lowest Tm of the pair ofprimers is usually used.

The final step of PCR amplification is the DNA extension from theprimers which is done by the thermostable Taq DNA polymerase usually at72° C., which is the optimal temperature for the polymerase to work. Thelength of the incubation at each temperature, the temperaturealterations and the number of cycles are controlled by a programmablethermal cycler. The analysis of the PCR products depends on the type ofPCR applied. If a conventional PCR is used, the PCR product is detectedusing for example agarose gel electrophoresis or other polymer gel likepolyacrylamide gels and ethidium bromide (or other nucleic acidstaining).

Conventional RT-PCR is a time-consuming technique with importantlimitations when compared to real time PCR techniques. This combinedwith the fact that ethidium bromide has low sensitivity, yields resultsthat are not always reliable. Moreover, there is an increasedcross-contamination risk of the samples since detection of the PCRproduct requires the post-amplification processing of the samples.Furthermore, the specificity of the assay is mainly determined by theprimers, which can give false-positive results. However, the mostimportant issue concerning conventional RT-PCR is the fact that it is asemi or even a low quantitative technique, where the amplicon can bevisualized only after the amplification ends.

Real time RT-PCR provides a method where the amplicons can be visualizedas the amplification progresses using a fluorescent reporter molecule.There are three major kinds of fluorescent reporters used in real timeRT-PCR, general nonspecific DNA Binding Dyes such as SYBR Green I,TaqMan Probes and Molecular Beacons (including Scorpions).

The real time PCR thermal cycler has a fluorescence detection threshold,below which it cannot discriminate the difference between amplificationgenerated signal and background noise. On the other hand, thefluorescence increases as the amplification progresses and theinstrument performs data acquisition during the annealing step of eachcycle. The number of amplicons will reach the detection baseline after aspecific cycle, which depends on the initial concentration of the targetDNA sequence. The cycle at which the instrument can discriminate theamplification generated fluorescence from the background noise is calledthe threshold cycle (Ct). The higher is the initial DNA concentration,the lower its Ct will be.

Other forms of nucleic acid detection can include next generationsequencing methods such as DNA SEQ or RNA SEQ using any known sequencingplatform including, but not limited to: Roche 454, Solexa GenomeAnalyzer, AB SOLiD, Illumina GA/HiSeq, Ion PGM, Mi Seq, among others(Liu et al., 2012 Journal of Biomedicine and Biotechnology Volume 2012ID 251364; Franca et al., 2002 Quarterly Reviews of Biophysics 35 pg.169-200; Mardis 2008 Genomics and Human Genetics vol. 9 pg. 387-402).

In other embodiments, nucleic acids may be detected with other highthroughput hybridization technologies including microarrays, gene chips,LNA probes, nanoStrings, and fluorescence polarization detection amongothers.

In some embodiments, detection of markers can be achieved at an earlystage of plant growth by harvesting a small tissue sample (e.g., branch,or leaf disk). This approach is preferable when working with largepopulations as it allows breeders to weed out undesirable progeny at anearly stage and conserve growth space and resources for progeny whichshow more promise. In some embodiments the detection of markers isautomated, such that the detection and storage of marker data is handledby a machine. Recent advances in robotics have also led to full serviceanalysis tools capable of handling nucleic acid/protein markerextractions, detection, storage and analysis.

Quantitative Trait Loci

Breeding schemes of the present application can include crosses betweendonor and recipient plants. In some embodiments, said donor plantscontain a gene or genes of interest which may confer the plant with adesirable phenotype. The recipient line can be an elite line havingcertain favorable traits for commercial production. In one embodiment,the elite line may contain other genes that also impart said line withthe desired phenotype. When crossed together, the donor and recipientplant may create a progeny plant with combined desirable loci which mayprovide quantitatively additive effect of a particular characteristic.In that case, QTL mapping can be involved to facilitate the breedingprocess.

A QTL (quantitative trait locus) mapping can be applied to determine theparts of the donor plant's genome conferring the desirable phenotype,and facilitate the breeding methods. Inheritance of quantitative traitsor polygenic inheritance refers to the inheritance of a phenotypiccharacteristic that varies in degree and can be attributed to theinteractions between two or more genes and their environment. Though notnecessarily genes themselves, quantitative trait loci (QTLs) arestretches of DNA that are closely linked to the genes that underlie thetrait in question. QTLs can be molecularly identified to help mapregions of the genome that contain genes involved in specifying aquantitative trait. This can be an early step in identifying andsequencing these genes.

Typically, QTLs underlie continuous traits (those traits that varycontinuously, e.g. yield, height, level of resistance to virus, etc.) asopposed to discrete traits (traits that have two or several charactervalues, e.g. smooth vs. wrinkled peas used by Mendel in hisexperiments). Moreover, a single phenotypic trait is usually determinedby many genes. Consequently, many QTLs are associated with a singletrait.

A quantitative trait locus (QTL) is a region of DNA that is associatedwith a particular phenotypic trait. Knowing the number of QTLs thatexplains variation in the phenotypic trait tells about the geneticarchitecture of a trait. It may tell that a trait is controlled by manygenes of small effect, or by a few genes of large effect or by a severalgenes of small effect and few genes of larger effect.

Another use of QTLs is to identify candidate genes underlying a trait.Once a region of DNA is identified as contributing to a phenotype, itcan be sequenced. The DNA sequence of any genes in this region can thenbe compared to a database of DNA for genes whose function is alreadyknown.

In a recent development, classical QTL analyses are combined with geneexpression profiling i.e. by DNA microarrays. Such expression QTLs(e-QTLs) describes cis- and trans-controlling elements for theexpression of often disease-associated genes. Observed epistatic effectshave been found beneficial to identify the gene responsible by across-validation of genes within the interacting loci with metabolicpathway and scientific literature databases.

QTL mapping is the statistical study of the alleles that occur in alocus and the phenotypes (physical forms or traits) that they produce(see, Meksem and Kahl, The handbook of plant genome mapping: genetic andphysical mapping, 2005, Wiley-VCH, ISBN 3527311165, 9783527311163).Because most traits of interest are governed by more than one gene,defining and studying the entire locus of genes related to a trait giveshope of understanding what effect the genotype of an individual mighthave in the real world.

Statistical analysis is required to demonstrate that different genesinteract with one another and to determine whether they produce asignificant effect on the phenotype. QTLs identify a particular regionof the genome as containing one or several genes, i.e. a cluster ofgenes that is associated with the trait being assayed or measured. Theyare shown as intervals across a chromosome, where the probability ofassociation is plotted for each marker used in the mapping experiment.

To begin, a set of genetic markers must be developed for the species inquestion. A marker is an identifiable region of variable DNA. Biologistsare interested in understanding the genetic basis of phenotypes(physical traits). The aim is to find a marker that is significantlymore likely to co-occur with the trait than expected by chance, that is,a marker that has a statistical association with the trait. Ideally,they would be able to find the specific gene or genes in question, butthis is a long and difficult undertaking. Instead, they can more readilyfind regions of DNA that are very close to the genes in question. When aQTL is found, it is often not the actual gene underlying the phenotypictrait, but rather a region of DNA that is closely linked with the gene.

For organisms whose genomes are known, one might now try to excludegenes in the identified region whose function is known with somecertainty not to be connected with the trait in question. If the genomeis not available, it may be an option to sequence the identified regionand determine the putative functions of genes by their similarity togenes with known function, usually in other genomes. This can be doneusing BLAST, an online tool that allows users to enter a primarysequence and search for similar sequences within the BLAST database ofgenes from various organisms.

Another interest of statistical geneticists using QTL mapping is todetermine the complexity of the genetic architecture underlying aphenotypic trait. For example, they may be interested in knowing whethera phenotype is shaped by many independent loci, or by a few loci, andhow those loci interact. This can provide information on how thephenotype may be evolving.

Molecular markers are used for the visualization of differences innucleic acid sequences. This visualization is possible due to DNA-DNAhybridization techniques (RFLP) and/or due to techniques using thepolymerase chain reaction (e.g. STS, SNPs, microsatellites, AFLP). Alldifferences between two parental genotypes will segregate in a mappingpopulation based on the cross of these parental genotypes. Thesegregation of the different markers may be compared and recombinationfrequencies can be calculated. The recombination frequencies ofmolecular markers on different chromosomes are generally 50%. Betweenmolecular markers located on the same chromosome the recombinationfrequency depends on the distance between the markers. A lowrecombination frequency usually corresponds to a low distance betweenmarkers on a chromosome. Comparing all recombination frequencies willresult in the most logical order of the molecular markers on thechromosomes. This most logical order can be depicted in a linkage map(Paterson, 1996, Genome Mapping in Plants. R. G. Landes, Austin). Agroup of adjacent or contiguous markers on the linkage map that isassociated to a reduced disease incidence and/or a reduced lesion growthrate pinpoints the position of a QTL.

The nucleic acid sequence of a QTL may be determined by methods known tothe skilled person. For instance, a nucleic acid sequence comprisingsaid QTL or a resistance-conferring part thereof may be isolated from adonor plant by fragmenting the genome of said plant and selecting thosefragments harboring one or more markers indicative of said QTL.Subsequently, or alternatively, the marker sequences (or parts thereof)indicative of said QTL may be used as (PCR) amplification primers, inorder to amplify a nucleic acid sequence comprising said QTL from agenomic nucleic acid sample or a genome fragment obtained from saidplant. The amplified sequence may then be purified in order to obtainthe isolated QTL. The nucleotide sequence of the QTL, and/or of anyadditional markers comprised therein, may then be obtained by standardsequencing methods.

One or more such QTLs associated with a desirable trait in a donor plantcan be transferred to a recipient plant to incorporate the desirabletrait into progeny plants by transferring and/or breeding methods.

In one embodiment, an advanced backcross QTL analysis (AB-QTL) is usedto discover the nucleotide sequence or the QTLs responsible for theresistance of a plant. Such method was proposed by Tanksley and Nelsonin 1996 (Tanksley and Nelson, 1996, Advanced backcross QTL analysis: amethod for simultaneous discovery and transfer of valuable QTL fromun-adapted germplasm into elite breeding lines. Theor Appl Genet92:191-203) as a new breeding method that integrates the process of QTLdiscovery with variety development, by simultaneously identifying andtransferring useful QTL alleles from un-adapted (e.g., land races, wildspecies) to elite germplasm, thus broadening the genetic diversityavailable for breeding. AB-QTL strategy was initially developed andtested in tomato, and has been adapted for use in other crops includingrice, maize, wheat, pepper, barley, and bean. Once favorable QTL allelesare detected, only a few additional marker-assisted generations arerequired to generate near isogenic lines (NILs) or introgression lines(ILs) that can be field tested in order to confirm the QTL effect andsubsequently used for variety development.

Isogenic lines in which favorable QTL alleles have been fixed can begenerated by systematic backcrossing and introgressing of marker-defineddonor segments in the recurrent parent background. These isogenic linesare referred to as near isogenic lines (NILs), introgression lines(ILs), backcross inbred lines (BILs), backcross recombinant inbred lines(BCRIL), recombinant chromosome substitution lines (RCSLs), chromosomesegment substitution lines (CSSLs), and stepped aligned inbredrecombinant strains (STAIRSs). An introgression line in plant molecularbiology is a line of a crop species that contains genetic materialderived from a similar species. ILs represent NILs with relatively largeaverage introgression length, while BILs and BCRILs are backcrosspopulations generally containing multiple donor introgressions per line.As used herein, the term “introgression lines or ILs” refers to plantlines containing a single marker defined homozygous donor segment, andthe term “pre-ILs” refers to lines which still contain multiplehomozygous and/or heterozygous donor segments.

To enhance the rate of progress of introgression breeding, a geneticinfrastructure of exotic libraries can be developed. Such an exoticlibrary comprises a set of introgression lines, each of which has asingle, possibly homozygous, marker-defined chromosomal segment thatoriginates from a donor exotic parent, in an otherwise homogenous elitegenetic background, so that the entire donor genome would be representedin a set of introgression lines. A collection of such introgressionlines is referred as libraries of introgression lines or IL libraries(ILLs). The lines of an ILL cover usually the complete genome of thedonor, or the part of interest. Introgression lines allow the study ofquantitative trait loci, but also the creation of new varieties byintroducing exotic traits. High resolution mapping of QTL using ILLsenable breeders to assess whether the effect on the phenotype is due toa single QTL or to several tightly linked QTL affecting the same trait.In addition, sub-ILs can be developed to discover molecular markerswhich are more tightly linked to the QTL of interest, which can be usedfor marker-assisted breeding (MAB). Multiple introgression lines can bedeveloped when the introgression of a single QTL is not sufficient toresult in a substantial improvement in agriculturally important traits(Gur and Zamir, Unused natural variation can lift yield barriers inplant breeding, 2004, PLoS Biol.; 2(10):e245).

Tissue Culture

As it is well known in the art, tissue culture of tomato can be used forthe in vitro regeneration of tomato plants. Tissues cultures of varioustissues of tomato and regeneration of plants therefrom are well knownand published. By way of example, a tissue culture comprising organs hasbeen used to produce regenerated plants as described in Girish-Chandelet al., Advances in Plant Sciences. 2000, 13: 1, 11-17, Costa et al.,Plant Cell Report. 2000, 19: 3327-332, Plastira et al., ActaHorticulturae. 1997, 447, 231-234, Zagorska et al., Plant Cell Report.1998, 17: 12 968-973, Asahura et al., Breeding Science. 1995, 45:455-459, Chen et al., Breeding Science. 1994, 44: 3, 257-262, Patil etal., Plant and Tissue and Organ Culture. 1994, 36: 2, 255-258. It isclear from the literature that the state of the art is such that thesemethods of obtaining plants are routinely used and have a very high rateof success. Thus, another aspect of this invention is to provide cellswhich upon growth and differentiation produce tomato plants having allthe physiological and morphological characteristics of hybrid tomatoplant HMX8148.

As used herein, the term “tissue culture” indicates a compositioncomprising isolated cells of the same or a different type or acollection of such cells organized into parts of a plant. Exemplarytypes of tissue cultures are protoplasts, calli, plant clumps, and plantcells that can generate tissue culture that are intact in plants orparts of plants, such as embryos, pollens, flowers, seeds, leaves,stems, roots, root tips, anthers, pistils, meristematic cells, axillarybuds, ovaries, seed coats, endosperms, hypocotyls, cotyledons and thelike. Means for preparing and maintaining plant tissue culture are wellknown in the art. By way of example, a tissue culture comprising organshas been used to produce regenerated plants. U.S. Pat. Nos. 5,959,185,5,973,234, and 5,977,445 describe certain techniques, the disclosures ofwhich are incorporated herein by reference.

EXAMPLES

The foregoing examples of the related art and limitations relatedtherewith are intended to be illustrative and not exclusive. Otherlimitations of the related art will become apparent to those of skill inthe art upon a reading of the specification.

Example 1—Development of New HMX8148 Tomato Variety

Breeding History of HMX8148

Hybrid tomato plant HMX8148 has superior characteristics. The female(DFTOMF3) and male (DFTOMM3) parents were crossed to produce hybrid (F1)seeds of HMX8148. The seeds of HMX8148 can be grown to produce hybridplants and parts thereof. The hybrid HMX8148 can be propagated by seedsproduced from crossing tomato inbred line DFTOMF3 with tomato inbredline DFTOMM3 or vegetatively.

The origin and breeding history of hybrid plant HMX8148 can besummarized as follows: the line DFTOMF3 was used as the female plant andcrossed by pollen from the line DFTOMM3 (both proprietary lines owned byHM.CLAUSE, Inc.). The first trial planting of this hybrid was done intwo different locations (Parrish and Immokalee) in Florida, UnitedStates in the fall and spring seasons of the first year of development.The hybrid was further trialed for three additional years, an example ofsuch trial being disclosed in Tables 2 and 3.

The inbred line DFTOMF3 is a parent with a medium-tall vine; thatproduces small, deep-round fruits. This inbred line was used as femaleparent in this cross.

The inbred DFTOMM3 is a parent with a small vine and very large fruit,with deep-round shape. It was used as the male parent in this cross.

Hybrid tomato plant HMX8148 is similar to hybrid tomato Southern Ripe, acommercial variety. As shown in tables 2 and 3, while similar to hybridtomato Southern Ripe, there are significant differences including theplant habit which is semi-erect for HMX8148 and erect for Southern Ripe.The fruit shape is round for HMX8148 and oblong for Southern Ripe. Theyalso differ in resistance to Tomato Yellow Leaf Curl virus with HMX8148having intermediate resistance while Southern Ripe is susceptible.

Some of the criteria used to select the hybrid HMX8148 as well as inbredparent lines in various generations include: plant vigor, fruit size,fruit firmness, fruit shape, and disease resistances.

Hybrid tomato plant HMX8148 has shown uniformity and stability for thetraits, within the limits of environmental influence for the traits asdescribed in the following Variety Descriptive Information. No varianttraits have been observed or are expected for important agronomicaltraits in tomato hybrid HMX8148.

Hybrid tomato plant HMX8148 has the following morphologic and othercharacteristics, as compared to SOUTHERN RIPE (based primarily on datacollected in Parrish, Fla., USA, all experiments done under the directsupervision of the applicant).

TABLE 1 Variety HMX8148 Southern Ripe Observation trial planted in:Field Field Observation trial planting type: Transplant Transplant Datesof seeding/transplanting: March/May March/May Observation trial plantingtype: Transplanted and Transplanted and staked staked Seedlinganthocyanin in hypocotyl of 2-15 cm: 1 = absent; 2 2 2 = present Matureplant: height growth type: 1 = indeterminate; 2 = determinate 2 2 form:1 = normal; 2 = compact; 3 = dwarf; 4 = brachytic 1 1 size of canopy(compared to others of similar form): 2 2 1 = small; 2 = medium; 3 =large; habit: 1 = sprawling; 2 = semi-erect; 3 = erect 2 3 branching: 1= sparse; 2 = intermediate; 3 = profuse 2 2 number of nodes betweeninflorescence 1 or 2 1 or 2 pubescence on younger stems: 1 = smooth (nolong 3 4 hairs); 2 = sparsely hairy (scattered long hairs); 3 =moderately hairy; 4 = densely hairy or wooly Leaf type: 1 = tomato; 2 =potato (Trip-L-Crop) 1 1 Morphology margins of major leaflets: 1 =absent; 2 = shallowly 2, 3 2 toothed or scalloped; 3 = deeply toothed orcut, specially towards base surface of major leaflets: 1 = smooth; 2 =rugose 2 2 (bumpy or veiny) pubescence: 1 = smooth (no long hairs); 2 =normal; 2 3 3 = hirsute; 4 = wooly Inflorescence type: 1 = simple; 2 =forked (2 major axes); 3 = 1 and 2 1 and 2 compound (much branched)number of flowers in inflorescence average 3 4 leafy or “running”inflorescence: 1 = absent; 2 = 2 2 occasional; 3 = frequent Flowercalyx: 1 = normal, lobes awl-shaped; 2 = macrocalyx, 1 1 lobes large, 3= fleshy calyx-lobes: 1 = shorter than corolla; 2 = approx., 1, 2, 3 1equaling corolla; 3 = distinctly longer than corolla corolla color: 1 =yellow: 2 = old gold; 3 = white or tan 1 1 style pubescence: 1 = absent;2 = sparse; 3 = dense 1 2 anthers: 1 = all fused into tube; 2 =separating into 2 or 1 1 more Fruit typical shape in longitudinalsection 3 = round 4 = oblong shape of transverse section: 1 = round; 2 =flattened; 1, 3 1, 3 3 = angular; 4 = irregular shape of stem end: 1 =flat; 2 = indented 2 2 shape of blossom end: 1 = indented; 2 = flat; 3 =2 2 nippled; 4 = tapered shape of pistil scar: 1 = dot; 2 = stellate; 3= linear; 2 2 4 = irregular abscission layer: 1 = present (pedicellate);2 = absent 1 1 (jointless) length of mature fruit (stem axis in mm) 65 78  diameter of fruit at widest point (mm) 65  79  number of locules: 1= two; 2 = three; 3 = four or five; 3 3 4 = more than 5 fruit base color(mature-green stage): 1 = light green; 3 3 2 = light gray-green; 3 =apple or medium green 4 = yellow green; 5 = dark green fruit pattern(mature-green stage): 1 = uniform 1 1 green; 2 = green-shouldered; 3 =radial stripes on sides of fruit fruit color full ripe: 1 = white; 2 =yellow; 3 = orange; 5 5 4 = pink; 5 = red; 6 = brownish; 7 = greenish; 8= other flesh color full ripe: 1 = yellow; 2 = pink; 3 = 3 3red/crimson; 4 = orange; 5 other flesh color: 1 = uniform; 2 = withlighter and darker 2 1 areas in walls locular gel color of table-ripefruit: 1 = green; 2 = 3 3 yellow; 3 = red ripening: 1 = inside out; 2 =uniformity; 3 = outside in 1 1 stem scar size: 1 = small (Roma); 2 =medium; 3 = 1 2 large core: 1 = coreless (absent or smaller than 6 × 6mm); 2 2 2 = present epidermis color: 1 = colorless; 2 = yellow 2 2epidermis: 1 = normal; 2 = easy-peel 1 1 field holding ability 4 out of5 4 out of 5 fruit harvestability: 1 = many rotten or broken; 2 = 4 4fruit soft, many rotten fruits; 3 = some rotten fruit; 4 = few rottenfruit; 5 = no rotten fruits, no rejected fruits Disease and pestreaction: 0 = not tested; 1 = highly resistant; 2 = resistant, fewsymptoms; 3 = resistance, few lesions in number and size; 4 = moderatelyresistance; 5 = intermediate resistance; 6 = moderate susceptible; 7 =susceptible; 9 = highly susceptible Virus diseases tobacco mosaic race 07 7 tobacco mosaic race 1 7 7 Tomato spotted wilt 7 5 Tomato yellow leafcurl 5 7 Fungal diseases Fusarium wilt race 1 (F. oxysporum f.lycopersici) 1 0 Fusarium wilt race 2 (F. oxysporum f. lycopersici) 1 2Fusarium wilt race 3 (F. oxysporum f. lycopersici) 1 2 Late blight, race0 (Phytophthora infestans) 7 7 Late blight, race 1 7 7 Verticillium wiltrace 1 (V. albo-atrum) 1 2 Grey leaf spot (Stemphylium spp.) 5 5 Insectsand Pests southern root knot nematode (M. incognia) 5 5 Other Maturity(in number of days to first harvest) 98  100  Fruit season(concentration): 1 = long; 2 = medium; 2 2 3 = short, concentrated; 4 =very concentrated Relative maturity in areas tested: 1 = early; 2 = 3 4medium early; 3 = medium; 4 = medium late; 5 = late; 6 = variableAdaptation Culture: 1 = field; 2 = greenhouse 1 1 Principle use(s): 1 =home garden; 2 = fresh market; 2 2 3 = whole-pack canning; 4 =concentrated products; 5 = multiuse; 6 = other Machine harvest: 1 = notadapted; 2 = adapted 1 1 Regions to which adaptation has beendemonstrated: 4 4 1 = Northeast; 2 = Mid Atlantic; 3 = Southeast; 4 =Florida; 5 = Great Plains; 6 = south central; 7 = Intermountain West; 8= Northwest; 9 = California (Sacramento and Upper San Joaquin Valley);10 = California (Coastal Areas); 11 = California (Southern San JoaquinValley & desserts); 12 = South American countries

Example 2—Comparison of New HMX8148 Tomato with Check Variety

In the tables that follow, the traits and characteristics of hybridtomato HMX8148 are given compared to another hybrid. The data collectedis presented for key characteristics and traits. Hybrid tomato HMX8148was tested at numerous locations. Information about the hybrid, ascompared to a check hybrid is presented (based primarily on datacollected in Florida, all experiments done under the direct supervisionof the applicant).

Tables 2 and 3 below show the characteristics of hybrid tomato HMX8148compared to hybrid Southern Ripe as measured in 2 growing seasons.Column 1 identifies the varieties, column 2 the fruit color with1=lightest; 9=darkest, column 3 the fruit shape with 1=flat round;2=medium round; 3=deep round, column 4 the fruit set with 1=lightest;9=heaviest, column 5 the plant height with 1=shortest; 9=tallest, column6 to 11 the % marketable fruit by diameter class with 4×4>9.3 cm;4×5>8.3 cm and <1=9.3 cm; 5×5>7.5 cm and <1=8.3 cm; 5×6>6.5 cm and<1=7.5 cm; 6×6>6 cm and <1=6.5 cm; 6×7>5.3 cm and </=6 cm. The 4×4, 4×5,5×5, 5×6, 6×6 and 6×7 is generally the number of rows and columns offruits that will fit into a standardized box. For example, a 4×4 meansthat 4 rows and 4 columns of fruits and therefore 16 tomato fruits willfit into the tomato box.

TABLE 2 Parrish, Florida season 1 % Marketable Fruit by Diameter ClassFruit Fruit Fruit Plant 4 × 4 × 5 × 5 × 6 × 6 × Variety Color Shape SetHeight 4 5 5 6 6 7 HMX8148 6 2, 3 7 4 0 1  7 16 45 31 Southern 5 2, 3 55 0 6 12 19 40 23 Ripe

TABLE 3 Parrish, Florida season 2 % Marketable Fruit by Diameter ClassFruit Fruit Fruit Plant 4 × 4 × 5 × 5 × 6 × 6 × Variety Color Shape SetHeight 4 5 5 6 6 7 HMX8148 6 2, 3 7 4 0 3  9 34 45  9 Southern 5 2, 3 55 0 4 21 50 15 10 Ripe

DEPOSIT INFORMATION

A deposit of the tomato seed of this invention is maintained byHM.CLAUSE, Inc. Davis Research Station, 9241 Mace Boulevard, Davis,Calif. 95618. In addition, a sample of the hybrid tomato seed of thisinvention has been deposited with the National Collections ofIndustrial, Food and Marine Bacteria (NCIMB), NCIMB Ltd. FergusonBuilding, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA Scotland.

To satisfy the enablement requirements of 35 U.S.C. 112, and to certifythat the deposit of the isolated strain of the present invention meetsthe criteria set forth in 37 CFR 1.801-1.809, Applicants hereby make thefollowing statements regarding the deposited hybrid tomato HMX8148(deposited as NCIMB Accession No. 43896).

1. During the pendency of this application, access to the invention willbe afforded to the Commissioner upon request;

2. All restrictions on availability to the public will be irrevocablyremoved upon granting of the patent under conditions specified in 37 CFR1.808;

3. The deposit will be maintained in a public repository for a period of30 years or 5 years after the last request or for the effective life ofthe patent, whichever is longer;

4. A test of the viability of the biological material at the time ofdeposit will be conducted by the public depository under 37 CFR 1.807;and

5. The deposit will be replaced if it should ever become unavailable.

Access to this deposit will be available during the pendency of thisapplication to persons determined by the Commissioner of Patents andTrademarks to be entitled thereto under 37 C.F.R. § 1.14 and 35 U.S.C. §122. Upon allowance of any claims in this application, all restrictionson the availability to the public of the variety will be irrevocablyremoved by affording access to a deposit of at least 2,500 seeds of thesame variety with the NCIMB.

INCORPORATION BY REFERENCE

All references, articles, publications, patents, patent publications,and patent applications cited herein are incorporated by reference intheir entireties for all purposes.

However, mention of any reference, article, publication, patent, patentpublication, and patent application cited herein is not, and should notbe taken as an acknowledgment or any form of suggestion that theyconstitute valid prior art or form part of the common general knowledgein any country in the world.

What is claimed is:
 1. A seed of hybrid tomato designated HMX8148,wherein a representative sample of seed of said hybrid has beendeposited under NCIMB No.
 43896. 2. A tomato plant, a part thereof, or acell thereof, produced by growing the seed of claim 1, wherein thetomato plant or a tomato plant regenerated from the part or the cell hasall the physiological and morphological characteristics of hybrid tomatodesignated HMX8148 when grown under the same environmental conditions.3. The tomato plant, the part thereof, or the cell thereof of claim 2,wherein the part is selected from the group consisting of a leaf, aflower, a fruit, a stalk, a root, a rootstock, a scion, a peduncle, astamen, an anther, a pistil, a pollen, an ovule, and a cell.
 4. A tissueculture of regenerable cells produced from the tomato plant or the partthereof of claim 2, wherein a tomato plant regenerated from the tissueculture has all the physiological and morphological characteristics ofhybrid tomato designated HMX8148 when grown in the same environmentalconditions.
 5. A tomato plant regenerated from the tissue culture ofclaim 4, said plant having all the physiological and morphologicalcharacteristics of hybrid tomato designated HMX8148 s when grown underthe same environmental conditions and wherein a representative sample ofseed of said hybrid has been deposited under NCIMB No.
 43896. 6. Atomato fruit produced from the plant of claim
 2. 7. A method forharvesting a tomato fruit, the method comprising: (a) growing the tomatoplant of claim 2 to produce a tomato fruit, and (b) harvesting saidtomato fruit.
 8. A tomato fruit produced by the method of claim
 7. 9. Amethod for producing a tomato seed, the method comprising: (a) crossinga first tomato plant with a second tomato plant and (b) harvesting theresultant tomato seed, wherein said first tomato plant and/or secondtomato plant is the tomato plant of claim
 2. 10. A method for producinga tomato seed, the method comprising: (a) self-pollinating the tomatoplant of claim 2 and (b) harvesting the resultant tomato seed.
 11. Amethod of vegetatively propagating the tomato plant of claim 2, themethod comprising: (a) collecting a part capable of being propagatedfrom the plant of claim 2 and (b) regenerating a plant from said part.12. The method of claim 11, further comprising (c) harvesting a fruitfrom said regenerated plant.
 13. A plant obtained from the method ofclaim 11, wherein said plant has all the physiological and morphologicalcharacteristics of hybrid tomato designated HMX8148 when grown under thesame environmental conditions.
 14. A fruit obtained from the method ofclaim
 12. 15. A method of producing a tomato plant obtained from hybridtomato designated HMX8148, the method comprising: (a) self-pollinatingthe tomato plant of claim 2 at least once to produce a progeny tomatoplant obtained from hybrid tomato designated HMX8148.
 16. The method ofclaim 15, further comprising the steps of: (b) crossing the progenytomato plant obtained from the hybrid tomato designated HMX8148 withitself or a second tomato plant to produce a progeny seed of asubsequent generation; (c) growing progeny plant from the progeny seedof the subsequent generation; (d) crossing the progeny plant of thesubsequent generation with itself or a second tomato plant to produce atomato plant derived from the hybrid tomato designated HMX8148; and (e)repeating step (b) and (c) for at least one generation to produce atomato plant further derived from the hybrid tomato designated HMX8148.17. A method of producing a tomato plant obtained from hybrid tomatodesignated HMX8148, the method comprising: (a) crossing the tomato plantof claim 2 with a second tomato plant to produce a progeny tomato plantobtained from hybrid tomato designated HMX8148.
 18. The method of claim17, further comprising the steps of: (b) crossing the progeny tomatoplant obtained from the hybrid tomato plant designated HMX8148 withitself or a second tomato plant to produce a progeny seed of asubsequent generation; (c) growing progeny plant from the progeny seedof the subsequent generation, (d) crossing the progeny plant of thesubsequent generation with itself or a second tomato plant to produce atomato plant derived from the tomato hybrid tomato plant designatedHMX8148; and (e) repeating step (b) and (c) to produce a tomato plantfurther derived from the hybrid tomato plant designated HMX8148.
 19. Amethod of producing a plant of hybrid tomato designated HMX8148comprising at least one desired trait, the method comprising introducinga single locus conversion conferring the desired trait into hybridtomato designated HMX8148 deposited under NCIMB No. 43896, whereby aplant of hybrid tomato designated HMX8148 comprising the desired traitis produced.
 20. A tomato plant, further comprising a single locusconversion and otherwise all of the physiological and morphologicalcharacteristics of hybrid tomato designated HMX8148 deposited underNCIMB No.
 43896. 21. The plant of claim 20, wherein the single locusconversion confers said plant with herbicide resistance.
 22. The plantof claim 20, wherein the single locus conversion is introduced into theplant by the use of recurrent selection, mutation breeding, wherein saidmutation breeding selects for a mutation that is spontaneous orartificially induced, backcrossing, pedigree breeding, haploid/doublehaploid production, marker-assisted selection, genetic transformation,genomic selection, Zinc finger nuclease (ZFN) technology,oligonucleotide directed mutagenesis, cisgenesis, intragenesis,RNA-dependent DNA methylation, agro-infiltration, TranscriptionActivation-Like Effector Nuclease (TALENs), CRISPR/Cas system,engineered meganuclease, engineered homing endonuclease, and DNA guidedgenome editing.
 23. A method of producing a tomato plant, comprisinggrafting a rootstock or a scion of the hybrid tomato plant of claim 2 toanother tomato plant.
 24. A method for producing nucleic acids, themethod comprising isolating nucleic acids from the plant of claim 2,part, or a cell thereof.
 25. A method for producing a second tomatoplant, the method comprising applying plant breeding techniques to theplant or part of claim 2 to produce the second tomato plant.